The impact of climate change on the symbiosis between dark septate endophytic fungi and rice plant

Plant roots play the main role in maintaining the resistant capacity to biotic and abiotic stress by forming a symbiotic relationship with endophytic fungi under [r]

VIETNAM NATIONAL UNIVERSITY, HANOI

VIETNAM JAPAN UNIVERSITY

MAI EI NGWE ZIN

THE IMPACT OF CLIMATE CHANGE ON THE SYMBIOSIS BETWEEN DARK SEPTATE ENDOPHYTIC FUNGI AND RICE PLANT

MASTER’S THESIS

VIETNAM NATIONAL UNIVERSITY, HANOI

VIETNAM JAPAN UNIVERSITY

MAI EI NGWE ZIN

THE IMPACT OF CLIMATE CHANGE ON THE SYMBIOSIS BETWEEN DARK SEPTATE ENDOPHYTIC FUNGI AND RICE PLANT

MAJOR: CLIMATE CHANGE AND DEVELOPMENT

CODE: 8900201.02QTD

RESEARCH SUPERVISORS:

PROF NARISAWA KAZUHIKO

DR HOANG THI THU DUYEN

Hanoi, 2020

PLEDGE

I assure that this thesis is my own research and has not been published The used of result of other research and other documents must comply with regulations The citations and references to documents, books, research papers and websites must be

in the list of references of the thesis

Author of thesis

MAI EI NGWE ZIN

TABLE OF CONTENTS

PLEDGE i

LIST OF TABLES iv

LIST OF FIGURES iv

LIST OF ABBREVIATIONS vi

ACKNOWLEDGMENT vii

FOREWORD viii

CHAPTER 1 INTRODUCTION 1

1.1 The Role of Rice Production in Asia 1

1.2 Vulnerable Rice Production under Climate Change (High Temperature) 3

1.3 The Concept of Endophytic fungi as Biofertilizers 4

1.4 Japonica Rice Species 6

1.5 Symbiosis Relationship between Plant and Fungi 7

1.5.1 Endophytic Fungi 7

1.6 The Impact of Climatic Change on the Symbiosis Relationship between Plant and Fungi 14

1.6.1 The Impact of temperature on the Soil Microbial Communities 14

1.6.2 The impact of Climate Factors on the Symbiosis between plant and Fungal Endophyte 15

1.6.3 The Effect of Seasonal Variation to Fungal Endophytes Communities 16

1.7 The Role of Fungal Endophytes in Sustainable Agriculture 17

1.7.1 Role of Fungal Endophytes in Controlling Environmental Contamination 18

1.7.2 The Role of Fungal Endophytes in Biotic and Abiotic stress resistance 18

1.7.3 The Role of Fungal Endophyte in Promoting the Plant Growth 21

1.7.4 Dark Septate Endophytic Fungi (DSE) and their abilities 22

1.8 Research Question and Hypothesis 24

1.9 Objective of the Research 24

CHAPTER 2 MATERIAL AND METHODS 27

2.1 Experiment Design 27

2.1.1 Research Parameter 27

2.2 Materials and Methods 27

2.2.1 Rice Seed Germination 27

2.2.2 Transplantation 29

2.2.3 The effect of high temperature on the symbiosis 30

2.3 The Second Experiment 31

2.4 Detection of DSE fungi in the root of the Koshihikari rice plant and identification of bacteria in the first experiment 31

2.4.1 Isolation and Identification of Bacteria 31

2.4.2 Checking the root colonization capacity of DSE endophyte in the roots of the Koshihikari rice plant 35

2.5 Identification of DSE fungi and Measurement of plant growth parameter of

2.5.1 Measurement of plant growth parameters 36 2.5.2 Identification of DSE fungi in the roots of the 10 days old Koshihikari Seedling 36

2.6 Statistical Analysis 37 CHAPTER 3 RESULTS AND DISCUSSION 39 3.1 Colonization capacity of DSE fungi in the root of the Koshihikari rice plant in the presence of bacteria 39

3.1.1 Identification of Bacteria from three isolates 39 3.1.2 Root colonization capacity of DSE Fungi in the roots of the Koshihikari rice plant in the presence of bacteria 41

3.2 Capacity of DSE fungi colonization and plant performance response to symbiosis under high-temperature treatment 44

3.2.1 Effect of DSE fungi symbiosis in plant growth 44 3.2.2 Identification of DSE fungi in the root of 10 days old Koshihikari rice seedling 45

3.3 The impact of high temperature on the symbiosis between DSE fungi and root of the Koshihikari rice plant 48 3.4 Proposed solutions for sustainable agriculture practice in the context of climate change 50

3.4.1 Recommendation for sustainable agriculture practice for Myanmar (future Research orientation) 51

REFERENCES 57 LISTS OF PUBLICATION BY THE AUTHOR 62

LIST OF TABLES

Table 1.1 Ideal temperatures for various development stages of the rice plant 3 Table 1.2 The Characterization of functional classes of endophytic fungi…………10 Table 2.1 Equipment and chemical used in rice seed germination 28 Table 2.2 Equipment and chemical used in transplantation 29 Table 2.3 Materials used in Bacteria DNA Extraction, PCR, and DNA sequencing 32 Table 2.4 Equipment used in Identification of DSE fungi 35

of the first experiment based on the NCBI BLAST database 39 Table 3.2 Result of the measurements of the physical parameters of the plant growth

Table 3.3 Flow Chart of Problem in Wet Tel Gu Groups of Villages, Myanmar 53

LIST OF FIGURES

Figure 1.1 The World’s Leading rice Producer during 2016-2019 1

Figure 2.1 Appearance of Bacteria Found in the first experiment during 24 hr of

transplantation 31

Figure 2.2 Isolation of single colony bacteria for DNA extraction 33

Figure 3.1 Colonization of Veronaeopsis simplex (Y34) DSE in the roots of 17 days

old Koshihikari rice seedling 42

Figure 3.2 Colonization of Cladophialophora chaetospira (OGR3) DSE in the roots

of 17 days old Koshihikari rice seedling 42

Figure 3.3 Colonization of Meliniomyces variabilis (J1PC1) DSE in the roots of 17

days old Koshihikari rice seedling showing an early developmental stage of an

intracellular microsclerotia (pointed with black arrow) and an early developmental

stage of an intracellular microsclerotia (pointed with white arrow) 42

old Koshihikari rice seedling 43

Figure 3.5 Colonization of Veronaeopsis simplex (Y34) DSE in the roots of 10 days

old Koshihikari rice seedling 46

of 10 days old Koshihikari rice seedling 47

Figure 3.7 Colonization of Meliniomyces variabilis (J1PC1) DSE in the roots of 10

days old Koshihikari rice seedling 47

Figure 3.8 Map of the Republic of the Union of Myanmar 51

LIST OF ABBREVIATIONS

ADB Asian Development Bank

AMF Arbuscular mycorrhizal fungi

BGA Blue Green Algae

BLAST Basic Local Alignment Search Tool

DSE Dark Septate Endophytic Fungi

FAO Food And Agriculture Organization

FAOSTAT Food and Agriculture Data

GCMs General Circulation Models

IPCC Intergovernmental Panel on Climate Change

IRRI International Rice Research Institute

J1PC1 Meliniomyces variabilis

LtPE2 Phialocephala fortinii

NCBI National Center for Biotechnology Information OGR3 Cladophialophora chaetospira

Y34 Veronaeopsis simplex

ACKNOWLEDGMENT

First and foremost, thanks to God, the Almighty, for His mercy and blessings throughout my two years of study of the master program including research work to complete the master’s program and research successfully

This research was implemented during my internship in Ibaraki University, Japan I

am grateful to Japan International Cooperation Agency (JICA) for their sponsorship during my internship under master program of Climate Change and Development, Vietnam-Japan University I am especially grateful to Prof Kazuhiko Narisawa for accepting me as his student and allowing me to conduct research in his laboratory under his supervision and sincere guidance I also would to express my appreciation

to Dr Duyen Thi Thu Hoang for her advice, support, and encouragement till the end

of my thesis

I would like to thank the laboratory members of the College of Agriculture, Ibaraki University, Japan, especially Ms Wiwiek Harsonowati from the Department of Symbiotic Science of Environment and Natural Resources, United Graduate School

of Agriculture Science, Tokyo University of Agriculture and Technology, Tokyo, Japan, for her help during the research

I also would like to express my gratitude to all members of the Department of Climate Change and Development, Vietnam Japan University, especially Dr Ahiko Kotera and Ms Bui Thi Hoa, as well as staffs from ICAS (Ibaraki University) as for their support and guidance during the entire my master program

Finally, a thank is not enough for my family, especially Miss Mai Ei Shwe Zin and

Mr Gabriel for their support to continue my study

FOREWORD

Rice production in South East Asia is under the threat of climate change such as

(IPCC AR5) In that case, endophytes can be used as the natural-based adaptation tools since they are found ubiquitously in different ecosystems However, symbiosis activity between the endophytes and their host plant under high temperature is not well known, requiring more elucidation The purpose of this research is to clarify the symbiotic features of endophytic fungi in the early stage of rice growth and to evaluate the impact of high temperature on rice-endophytic symbiosis through the observation of the fungi present in the roots The symbiosis activity between the dark septate endophytes (DSE) and Koshihikari rice plant under high temperature was

tested by comparing morphophysiology of rice colonize with Cladophialophora

chaetospira (OGR3), Meliniomyces variabilis (J1PC1), Phialocephala fortinii (LtPE2), Veronaeopsis simplex (Y34) and without colonization under continuous high

the plant growth parameters such as root/shoot ratio, shoot high, etc., were measured The colonization capacity of DSE fungi in the roots of the rice plant was demonstrated for all the selected species at 35ºC The research will provide fundamental scientific background of endophytic fungi application for global warming application in agriculture

CHAPTER 1 INTRODUCTION

1.1 The Role of Rice Production in Asia

More than 2.2 billion individuals in Asia depend on agribusiness (Asia Development Bank 2009) and 90 percent of the world's paddy cultivation and utilization are from Asia Rice production in the monsoon land that is known for Asia begins from the Islamic Republic of Pakistan to Japan, where the populace is densely conveyed and

a significant consumer of rice in the world China and India are recorded as the world's biggest rice producers by a wide margin The harvested region in India is bigger than China, however, the yield in China is higher than India since about all of the harvested area in China is irrigated, while practically 50% of India's rice zone is inundated Southeast Asia nations, for example, Indonesia, Bangladesh, Vietnam, Myanmar, and Thailand trails China and India During 2008-2010, the normal rice production from these seven nations was in excess of 30 billion tons of paddy according to IRRI 2013 Furthermore, as indicated by the source from the United States Department of Agriculture, China was driving as the world's first rice producer

At that point, India, Indonesia, and Bangladesh followed in the year 2016/2017 to 2018/2019

Figure 1.1 The World’s Leading rice Producer during 2016-2019 (source: United

States Department of Agriculture, 2019) India, Thailand, and the Socialist Republic of Viet Nam are the significant exporter

of rice on the world and possess in excess of 60% of the world's rice market during

the most recent decade (FAO, 2018) As indicated by India involved the most elevated fare volume of rice for around the world, by 12.5 million metric tons starting

in 2018/2019 Thailand followed the second-biggest rice exporter, with about 10.3 million metric tons of rice worldwide in that year Concerning the Southeast Asia nations, Vietnam, Burma (The Republic of the Union of Myanmar) and Cambodia were among the world's driving rice trading nations with 7, 3, and 1.3 million metric tons for the year 2018/2019 respectively

Even though China is the world's greatest cultivation and consumer country of rice, from the year 2015/2016 to 2017/2018, according to the United States Department of Agriculture, 2019, China turned into the top rice bringing in the nation over the world

As per the "Exploration Report on Paddy and Rice Import in China, 2019-2023", Dublin, Jan 18, 2019 (GLOBE NEWSWIRE), the explanation might be that because

of the quick urbanization and monetary development, many work power from the rural area moved to the modern segment since horticultural part contributes little pay

in China, which lead to slow-moving in agrarian yield and because of the significant expense of What's more, the subsequent factor is that the cost of domestic rice is higher than imported rice China's rice imports are predominantly from Vietnam and Thailand Their costs are just 80% of those of locally developed rice at a similar quality level or lower as indicated by the examination report on paddy and rice import

in China, 2019-2023

For Southeast Asia, thirty-one percent of the global rice harvest was from the region (FAOSTAT 2012) and is anticipated to represent 53% of net fares, 14% of net imports, 29% of the harvested territory, 25% of total production, and 22% of all-out rice consumption for the worldwide rice showcase throughout the following decade (ADB, 2012) Rainfed rice production system occupied 55 percent of paddy cultivation in Southeast Asia Therefore, the precipitation pattern plays a vital role in paddy cultivation of Southeast Asia Meanwhile, Asian Monsoon is the characteristic

of Asia, the biggest landmass on the world and it is encircled by the Arctic, the Indian, and the Pacific Oceans prompting ocean land warm difference and described by Asian Monsoon climate The dispersion of water assets in Asia is lopsided and prompts flooding and drought much of the time because of strong horizontal and vertical

gradient in both temperature and precipitation (Yanai and Li 1994, Qiu 2008, Yao et

al 2012, Sharma et al 2016) As indicated by the IPCC AR5 report, in tropical and

late-twentieth-century levels will negatively affect the significant yield production without adjustment Besides, as per the most models, utilizing a scope of General Circulation Models (GCMs) and Special Report on Emission Scenarios (SRES) situations, high temperature will prompt the lower measure of rice production because of the shorter developing time frame

1.2 Vulnerable Rice Production under Climate Change (High Temperature)

Rice is delicate to extreme climate stress, for example, high temperature will affect the basic development stages, floods cause halfway or complete submergence, additionally, it is exceptionally sensitive to salinity identified with sea-level rise and dry season spells by rain-fed paddy Rice is ordinarily developing during mid-year (summer) in tropical and sub-tropical locales and heat stress is a typical compel during anthesis and grain-filling stages (Kobata and Uemuki, 2004) The ideal temperatures for various development and advancement phases of rice are as follow: Table 1.1 Ideal temperatures for various development stages of the rice plant

The fitting temperature for typical advancement of rice is between 27 to 32ºC and higher temperature over 32 ºC will influence practically all phases in the development

of rice, Tian et al Temperature of rice-growing regions has already reached the optimal temperature of rice growth and high temperature is known to upset water, irons, and natural solute development across plant films, which meddles with

photosynthesis and plant respiration Halford, 2009 reported that when the plant is under prolonged high-temperature, electrolytic leakage may occur from leaves While another examination expressed that temperatures past the basic limits will decrease the development length of the rice crop just as expanded the spikelet sterility, diminishing grain-filling term, and upgrading respiratory losses, prompting lower yield and lower quality rice grain (Fitzgerald and Resurreccion 2009, Kim et al., 2011) It is projected that worldwide rice production, grain quality, and healthful advantages will be seriously diminished by increasing mean air temperature, (Teixeira et al., 2013, Lin et al., 2010, Wang et al., 2011)

According to IPCC AR4, it is projected that the world’s temperature will be increased

1ºC over the basic temperature (>24 ºC) will prompt a 10% decrease in both grain yield and biomass (S Mohanty, R Wassmann, A Nelson, P Moya, and S.V.K Jagadish, 2012) Moreover, IPCC anticipated that that "the quantities of cold days and nights have diminished and the quantities of warm days and nights have expanded across the majority of Asia since around 1950, and heatwave recurrence has expanded since the center of the twentieth century in enormous pieces of Asia" (IPCCAR5, Chapter 24) Endophytes as biofertilizers can be used as one of the natural-based adaptation tools since they are found obviously in different ecosystems

1.3 The Concept of Endophytic fungi as Biofertilizers

Agriculture is one of the most vulnerable sectors under climate change, which in turn contributes to climate change through land degradation, methane production, and so

on In Asia, extreme use of chemical fertilizer reasons soil pollution, nutrient imbalance, and soil erosion and that they lead to unfavorable effects on soil fertility (Zhang et al., 1996 and Hedlund et al., 2003) Therefore, the utility of biofertilizers promising in sustainable and ecofriendly agriculture cultivation while ensuring food demand for the world population

There are various varieties of biofertilizer including:

(a) Nitrogen-fixing biofertilizer: (i) symbiotic nitrogen fixer, (ii) symbiotic nitrogen fixer

non-(b) Phosphorous mobilizing biofertilizers: (i) Phosphate solubilizer, (ii) Phosphate absorber

(c) Organic matter decomposer biofertilizers: (i) Cellylolytic, (ii) Lignolytic All those styles of biofertilizers may be bacteria, fungi, and cyanobacteria

Nitrogen is one of the important nutrients for plant life and crop yields because it's far a primary component of chlorophyll, by which plants utilize sunlight and convert sugars from water and carbon dioxide from the air called photosynthesis Moreover, Nitrogen is also a basic component of amino acids, which are building blocks of proteins Although nitrogen is abundant in the air, plants cannot use the gaseous form

fixation The nitrogen fixation can occur in 3 ways:

Biologically- organisms in the soil convert the gaseous state of nitrogen into

Industrially- by applying nitrogen fertilizer

Nitrogen fixation can be also formed through symbiosis relationships between soil microbes and plants In the process, bacteria colonize plant roots, they reside and multiply After that, the association of plant and bacteria synchronize and stimulates plant growth processes such as the formation of root nodules in which they convert free nitrogen to ammonia, enlargements of plant cells, etc Blue-green algae (BGA) called cyanobacteria can be found on land and in water They also fix nitrogen and

are used as biofertilizers for cereal crops and non-legumes Oscillatoria, Nostoc, and

Anabaena, etc are examples of cyanobacteria The symbiotic association between the

aquatic fern Azolla and Anabaena is very important for paddy and in this microbial association, Anabaena receives carbon and nitrogen from the plant in exchange for fixed

nitrogen

Phosphorous is one of the crucial nutrients for plant growth, regulating plant physiology such as expanding stalk and stem quality, improve blossom arrangement, and expanding the nitrogen N-fixing limit of vegetables and protection from plant

infections By which phosphorous supports the growth of the whole life cycle of the plant The symbiosis association can also be found between plant and fungi and are classified as "mycorrhiza" The fungi affiliation serves the host plant by retaining the phosphorus from the soil and bolster the host plant Also, through the fungi affiliation, the host plant can resilience to biotic and abiotic stress, protection from the pathogen, and generally increment the plant development

It was reported that soil-infertility is the major constrain for low productivity especially in developing countries, Khosro and Yousef, 2012 Therefore, to maintain

biofertilizers enhances soil particles and continues soil fruitfulness and upgrades root expansion by producing the plant growth-promoting hormones (Subba Rao, 1993) Application of biofertilizers is known to improve soil fruitfulness and harvest efficiency in several crops, for example, improving the rice production by applying the high yielding assortments and through advancing the agrarian practices, for example, organic manures and biofertilizers application (Zaki et al., 2009) High yield

of different vegetables after immunization with nitrogen-fixing microorganism allow fixed nitrogen to plants as well as the Nitrogen status of soil (Zaidi et al., 2003) Field execution of excrement from biogas plants has been found to builds the yield just as soil microbial activity in maize (Mehetre and Kale, 2007) Biofertilizers are fit for providing significant supplements as well as micronutrients for the successful development of plants through the association of microorganisms

1.4 Japonica Rice Species

Two varieties of rice can be found in Asia, they are Japonica and Indica Japonica species can be grown in temperate upland regions of East Asia, Southeast Asia, and high elevations in South Asia where Indica can be grown for the most part in the marsh and lowered, all through tropical Asia Japonica rice grains have rounder, thicker in appearance, and harder qualities, contrasted with longer, more slender, and fluffier Indica rice grains Also, they are stickier because of the higher content of amylopectin Japonica species can be divided into three subgroups: 1) temperate Japonica, 2) tropical Japonica, and 3) aromatic Koshihikari, the famous cultivar of Japonica rice and bread in 1956 It is the most popular cultivar in Japan and occupied

as the most cultivation area in Japan since 1979 Koshihikari has a highly adaptive

capacity to the climate that they can be cultivated at altitudes ranging from 40°N to

31°N The vegetative phase of Koshihikari was 35.2 days if they are grown under

short-day conditions after sprouting at 28 °C (Mimoto et al 1989) and are highly

sensitive to temperature (Hosoi 1979) Meanwhile, tropical Japonica can be found in

the Central Dry Zone, Delta Region, and Coastal Region of Myanmar (21.9162° N,

95.9560° E), Ohm Saw, Genetic Diversity of Myanmar rice Cultivar The global

mean temperature has risen and the meteorological research projects an increase of

one or two degrees all over Japan in 2081 to 2100, which may cause threats to rice

productivity and growth Therefore, the symbiosis relationship with the fungal

endophyte in rice roots is promising to enhance the resistant capacity of plants in

coping with water stress and soil nutrient deficiency due to climate change impacts

1.5 Symbiosis Relationship between Plant and Fungi

Symbiosis: “Any association between two species populations that live together is

relationship microbes including mycorrhizal fungi and fungal endophytic (R

J.Rodriguez, et al, 2009 and Petrini, 1986) Therefore, mycorrhizas and fungal

endophytes are the two main groups of symbiotic association with plants and their

symbiosis relationship impact the structure and function of the ecosystem (Smith SE

and Read, 1997)

1.5.1 Endophytic Fungi

Endophytic fungi definition is that "endophytic fungi live within the tissue of the

plant and do not cause any symptoms of disease" (Sumita Jha, 2019) Fungal

endophytes can be discovered from all species of plants including temperate and

tropical forest, grassland and savannas as well as croplands, Mishra, Y., Singh, A et

al., 2014 The endophytic fungi colonize the host plant by their hype and infiltrate the

plant tissue, for example, through contaminated structures, such as appressoria (Ernst

et al., 2003) and develop intercellularly with virtually no cell injury (Rodriguez et

al.,2008a) The endophytic fungi can be isolated from the different parts of host plants

such as rots, stem as they reside entirely in the plant tissue and can be grown in the

artificial medium Endophytic fungi have huge biodiversity, particularly in the tropical and temperate rainforest Unlike AMF, the endophytic fungi colonize only through the root of the host, and live inside roots, stems, leaves, and develop during host senescence (Saikkonen et al., 1998) Their symbiosis benefit is that endophytic fungi receive protection and nutrients from the host, whereas the host plants may benefit from the endophytic fungi through increased resistance to biotic stress such

as herbivores and pathogens, and also various abiotic stresses and enhanced competitive abilities (Wilson, 1995; Rodriguez and Redman, 1997; Lehtonen et al., 2005)

1.5.1.1 Endophytic Fungi Classification and host range

Fungal endophytes can be classified into two basic groups according to their taxonomy, host range, colonization transmission design, tissue characteristics and ecological function, clavic-endophytes (C-endophytes), which are connected with grasses and non-Clavic-endophytes (NC-endophytes) Table 1.4 describes the characterization of the functional class of four classes of endophytic fungi

Class I Clavicipitaceous Endophytes: these are a relatively modest number of species that are limited to a few monocot hosts and contain animal-harming alkaloids The Clavicipitaceous endophytes symbiosis with grasses was first noticed by European specialists (Guerin, 1898; Vogl, 1898) Bacon et al., (1977) responded with the hypothesis that animals that consume contaminated tissue from endophytic fungi are related to the poisonous disorder Clavicipitaceous Endophytes have many benefits

in hosting plants such as:

Insect deterrence: insect feeding resistance of mushrooms to plants through the release of loline and peramine-mychotoxin alkaloidis (Clay 1990; Patterson et al 1991) has been observed to resist insect feeding to host plants (Rowan and Gaynor 1986)

Prevention of mammalian herbivores Some Clavicipitaceous Endophytes have a resilience from mammalian herbivores feeding to plants due to their releasing of mycotoxins, like ergot and lolithrem alkaloids (White, 1987 and Gentile et al.)

Nematode decrease: Endophytic fungus Acremonium coenophialum is described to

be resistant to infection by decreasing the number of nematode populations in fields

of Festuca arundinacea by an infection in high fescus (Kimmons et al., 1990)

Increased host disease resistance: Certain Clavicipitaceous Endophytes increase their

resistance to host plant disease The release, in symbioses’ with Epichloë festucae

(cool-seaon-grease), 2010 of the indole supplies, such as the secretion of sesquiterpene and the diacetamide compound, will resist the growth of other pathogenic fungi

Boost the ecological physiology of host plants: Arechavaleta et al., 1989 have indicated that some Clavicipitaceous Endophytes can withstand abiotic stress, such

as drought and contamination by chemicals The host plant can be encouraged by certain endophytes to extend the hair of the root, through which 'phenolic compounds' fill the rhizo The host plant will assimilate higher quantities of soil phosphorus and improve tolerance to aluminum by chelation according to this mechanism (Malinowski and Belesky 2000)

Class II Nonclavicipitaceous Endophytes (NC Endophytes): According to Rodriguez

et al., (2009), the class II NC-endophytes represent three distinct functional groups The host range of NC endophyts is very wide and includes both monocots and dicots (Rodriguez and Redman, 2004) and tropical leaves (Lodge et al., 1996; Frưhlich and Hyde, 1999; Arnold et al., 2000; Gamboa and Bayman, 2001) In biomes ranging from tropically woodland to boreal including Arctic-Antarctic communities (Carroll

& Carroll, 1978; Petrini, 1986; Stone, 1988), they can grow in both the above and lower ground tissues of the non-vascular plants, seedless vascular plants, conifers, woody, and herbaceous angiosperms And it looks like tolerance to stress adapted to the climate (Rodriguez et al 2008) Symbiosis with class II NC endophytes has many advantages for hosting plants:

Avoiding abiotic stress: The host may tolerate environmental stress and are typical for the high frequencies of infection of plants that are developing in a high natural stress setting, depending on its ability to colonize asymptomatically and confer

habitat-adjusted habitat (Redman et al 1999a, 2001, 2002; Rodriguez and Redman 2007)

Increase of biomass: Tudzynski and Sharon (2002) expressed that class II NC endophytes are able to develop shoot and additionally root biomass because of the enlistment of plant hormones by the host or through biosynthesis of plant hormones

by the fungi

Protection from fungal pathogen: Class 2 NC endophytes can resist fungal pathogens

by their various strategies including the production of secondary metabolites (Danielsen et Jensen, 1999; Narisawa et al., 2002; Campanile et al., 2007) (Schulz et al., 1999)

Table 1.2 The Characterization of functional classes of endophytic fungi

Endophytes

Non- Clavicipitaceous Endophytes

Fitness benefits Non-habitat adapted Non-habitat

adapted and habitat-adapted

habitat adapted

habitat adapted

Non-1.5.1.2 Bioactive compounds produced by fungal endophytes

Protein synthesis and respiration associated fungal metabolites are diverse, and several secondary metabolites have been isolated frequently and are chemically defined, which include waste products as well as pigments, toxins, and antibiotics, and have biological functions Schulz et al., 2002 reported that at least a research organism of antibacterial, fungicidal, algicide, and herbicide activities comprised 83% of the algal isolates and 80% of the endophytic fungi isolated from plants and only 64% of soil operation

Diverse classes of natural compounds produced by Fungal Endophytes

Alkaloids: A chemical compound containing basic nitrogen atoms and produced as common secondary metabolites by fungal endophytes occurs naturally The endophytic fungal-producing alkaloids have antimicrobial activity (Souza et al., 2004) Chaetomium globosum fungal endophyte isolated from Ginkgo biloba and the species produces chaetoglobosins A, G, V, Vb, and C, some of which have been shown to be cytototoxic (Li et al., 2014) Vincristine is an alkaloid from Catharanthus

roseus originally isolated from vinca (Zhang et al., 2000) Chaetoglobosin U is an alkaloid dependent on cytochalasin, isolated from Chaetomium globosum sp of the stem of Imperata cylindrica (Ding et al., 2006)

Phenols: Some fungal endophyte may isolate the phenols and phnlolic acids (Yu et

al., 2010) The endophytic Cytonaema sp fungi isolated from Quercus sp., produce

the isomeric novel tridepsides cytonic acids A and B which are identified as protease

inhibitors of the human cytomegalovirus (Guo et al., 2000) Nodulisporium sp endophytic fungi isolated from Juniperus cedar at Gomera Island which produces

two antimicrobial flavonoids (Dai et al., 2006)

Steroids: Fungal endophytes produce steroids and most of the isolated compounds have moderate antimicrobial activities The liquid culture of fungal endophyte

Colletotrichum sp isolated from Artemisia annua produce

3β-hydroxy-5α,8α-epidioxyergosta-6,9,22-triene and 3-oxoergosta-4-ene, two new steroids,

acetoxyergosta-7,22-diene along with ergosterol Some of them have antifungal

activity and have resistant to Gaeumannomyces graminis var tritici, Rhizoctonia

cerealis, Helminthosporium sativum, and Phytophthora capisici (Lu et al., 2000; and

Yu et al., 2010)

Terpenoids: Fungal endophytes produce large terpenoids such as Sesquiterpenes, Diterpenoids, and Triterpenoids (Yu et al., 2010) Sixty-five sesquiterpenes, forty-five diterpenes, five monoterpenes and twelve other terpenes were classified as isolates from fungal endophytes, out of a total of 127 terpenoid compounds, all of which have anti-microbial, anti-cancer and anti-protozoan activity (Souza et al., 2011)

Quinones: Some fungal endophytes produce quinones that significantly inhibit

pathogenic development Edenia gomzpompae, the endophytic fungi that contain quinones such as spiroketals (Wiyakrutta et al., 2004) The Pestalotiopsis microspora endophytic fungi, isolated from Torreya taxifolia, develop Torreyanic acid which is

an unusual dimeric quinone (Lee et al., 1996) Hormonema dematioides, fungal endophyte isolated from balsam fir produce rugulosin with an insecticide (Findlay

et al 1997) Coniothyrium sp the endophytic fungus, produce the new ras

farnesyl-protein transferase inhibitors Preussomerin N1, palmarumycin CP4a and palmarumycin CP5 ((Tan and Zou, 2001)

Peptides: Endophytic fungi develop peptides and some have significant antimicrobial

activity Acremonium sp endophyte sp Produce di-O-b-glucoside leucinostatin A and

leucinostatin A when grown in liquid culture (Strobel et al., 1997a and Kharwar et

al., 2011) The endophyte fungal Aspergillus rugulosus, and A Varies nidulans

Echinulate has an antimicrobial activity Cryptosporiopsis cf quercina endophytic

fungi isolated from Redwood produces cryptocandin, which has an antifungal activity (Tan and Zou, 2001)

Polyketides: Fungal endophyte Periconia sp F-31 isolated from the medicinal plant

Annona muricata, produce a new polyketide synthase-nonribosomal peptide

synthetase hybrid pericoannosin B (Zhang et al, 2016) A tryptophan−polyketide hybrid, Codinaeopsin, isolated from the fungal endophyte CR127A that was

symbiosis with a white yemeri tree Vochysia Guatemalensis in Costa Rica, which has the ability against Plasmodium falciparum, the causative agent of the most lethal form

of malaria (Kontnik and Clardy, 2008) Endophytic fungi Chaetomium globosum,

which was found in the leaves of Ginkgo biloba produce chaetoglobosins A and C, which were isolated from the EtOAc These compounds have the activity against the

growth of brine shrimp Artemia salina and Mucor miehei (Qin et al., 2009) The three

polyoxygenated polyketides, epicolactone, epicoccolides A and B were shown to have antimicrobial activities and significant inhibitory effects on the mycelia growth

of two peronosporomycete phytopathogens such as Pythium ultimum and Aphanomyces cochlioides, and the basidiomycetous fungus Rhizoctonia solani, were

isolated from the fungal endophyte of Epicoccum sp CAFTBO, symbiosis with

Theobroma cacao (Talontsi et al., 2013)

Acids: The fungal endophyte Salvia miltiorrhiza produces salvianolic acid C (Li et

al., 2016) New acid of pentamethyl- hexacosa-4E,8E,12E,16,18-pentaenoic acid were isolated from the endophytic fungi Phoma glomerata D14 (Khiralla ,2015) The fungal endophyte

3,7,11,15-Tetrahydroxy-18-hydroxymethyl-14,16,20,22,24-Curvularia papendorfii symbiosis with Vernonia amygdalina produce Khair acid,

which has antibacterial effect against the methicillin-resistant to Staphylococcus aureus with MIC value of 62.5 μg mL-1 The three endophytic fungi: Fusarium solani,

F oxysporum and F proliferatum isolated from pigeon pea Cajanus cajan they can produce cajaninstilbene acid (CSA), which is one of the major stilbenes found in pigeon pea The acid is revealed as hypotriglycerimic, hypoglycemic, anti-inflammatory, analgesic and antioxidant activities (Zhao et al., 2012) The solid-state

fermentation of the endophytic fungi Cytonaema sp produce Cytonic acids A and B

which have potential anti-viral effect against human cytomegalovirus (hCMV)

protease with values of IC50 = 43µM and IC50 =11µM respectively (Guo et al.,

2000) The cryptocin was isolated from Cryptosporiopsis cf quercinain, which has

antimycotic activity against several plant pathogenic fungal strains including Pythium ultimum, Pyricularia oryzae, with MIC values 0.39 and 0.78 μg mL-1 respectively (Li et al., 2000)

1.6 The Impact of Climatic Change on the Symbiosis Relationship between Plant and Fungi

1.6.1 The Impact of temperature on the Soil Microbial Communities

Temperature influenced the physical and biochemical mechanisms of soil as well as regulating the soil microbial activities The increased surface temperature corresponds with decreased soil moisture, Venkat Lakshmi et al., 2003 Soil moisture controls the soil microorganism as well as biochemical activities of soil (A Borowik and J Wyszkowska, 2016) It is due to the fact that soil microorganism can rapidly equilibrate to the osmotic conditions of their environment and remain hydrated when the soil near them are dried by accumulating solutes to retain water within their cells

the substrates of microbes by dissolution, diffusion, and transportation

erode hillsides and scour stream channels Therefore water always flows downhill due to the gravitational potential gradient And in the process of osmosis, water on the freshwater side of a membrane moves across to the salty side because the solute potential on the salty side is lower The total water potential gradient made water to transfer from soil through plants to the atmosphere

For the soil microbes, water potential is a basic in controlling their endurance and capacity since they are in micro size and cozy contact with their environment and are along these lines essentially equilibrate with the water potential in the soil around them At the point when the soil got dried, the total water potential drops and the organisms must gather solutes to bring down their inside solute potential to coordinate the water capability of the encompassing soil to abstain from losing water to their condition and drying out When the cellular water potential of microbes drops, it might prompt a loss of cell turgor (Harris 1981), practically equivalent to shrinking

in plants This process interferes with microbial physiological capacities and prompts decreasing the metabolic rates and this can even lead their cell capacities to fall flat, slaughtering the microbes

The global mean surface temperature has risen during the previous century (Jones et

al 2012, Stocker et al 2013, Sun et al 2017) and according to IPCC AR5, it is projected that heat waves will prone to be progressively extreme, successive and cold scenes are anticipated in a diminishing later on The daily minimum temperature is projected to increase than the daily maximum temperature

1.6.2 The impact of Climate Factors on the Symbiosis between plant and Fungal Endophyte

Previous research has argued that climate factors, for example, precipitation and atmospheric humidity may have an impact on the event of some endophytic species, Petrini, 1991 In this research, the foliar endophytic fungal communities of the species Musa acuminate (Banana) were investigated Sampling was taken from Hong Kong, which represented a location outside the natural range of Musa acuminate (Banana), and Queensland, Australia, which represented part of the endemic range in order to test whether host plants outside their natural range supported different endophytic assemblies to host plants within the natural range As a result, there was a high species

abundance in the southeastern Queensland plantation The researcher argued that

"this variation is probably a reflection of the range of environmental conditions sampled in Hong Kong and northern Queensland Sites differ primarily from rainfall, altitude, and forest types; these are all factors that may influence the abundance of fungal species as they affect moisture, temperature, and potential inoculum sources” The impact of climate on symbiotic fungal endophyte diversity and performance was further explained by Hannah Qiauque and Christine V Hawkes, 2013 In this research, endophytes in grasses across the steep precipitation gradient of Edwards Plateau in Central Texas were collected to elucidate the relative importance of environmental and spatial factors in the structuring of endophyte communities Annual rainfall across the Plateau varies from ~40 to 90 km west to east, with an average annual precipitation change of ~10 cm every 40–50 km The 20 endophyte isolates in symbiosis with grass seedlings were also sampled from drier and wetter regions with high and low soil moisture in the greenhouse Environmental factors related to historical and current precipitation have been the most important predictors

of endophyte communities in the field due to the possibility of past drought patterns that create legacies that constrain the current community and ecosystem properties The historical mean annual and current spring rainfall also affected around 35% of the variation in the endophyte community composition For the performance of fungal endophytic fungi, the reduction of plant water loss in the greenhouse was reduced for endophytic fungi in western sites compared to fungi in eastern sites They concluded that although historical and current climatic factors have an impact on the endophyte community, their symbiotic function could not be predicted by the local environmental condition

1.6.3 The Effect of Seasonal Variation to Fungal Endophytes Communities

The impact of seasonal variation and environmental factors on endophytes transmission in tall fescue was studied by Ho J Ju, 2003 In this study, the impact of seasonal variation on endophytes in tall fescue was investigated by collecting the

Watkinsville, GA, and Oregon And the result showed that endophyte frequency was greater when sampled from April to December in Georgia and from October through

January to May in Oregon The temperature impact was tested by Jesup Max Q seeds

in the greenhouse and four temperature treatment regimen were established: (1) 12/6

° C day/night temperature for 3 weeks, 25/19 ° C day/night for 3 weeks, (2) 25/19 °

C day/night temperature for 3 weeks, 12/6 ° C day/night for 3 weeks, (3) 12/6 ° C day /night temperature for 6 weeks, and (4) 25/19 ° C day /night temperature for 6 weeks And it has resulted that plants grown at higher temperature regimes showed higher growth and greatest dry matter throughout the experiment than those grown at the cooler temperature regime And the endophyte concentration was increased at the 25/19ºC regime, throughout the experiment

J Collodo et al., 1999 investigated the impact of geographical and seasonal influences

on the distribution of fungal endophytes, Quercus ilex samples were collected from

four sites in central Spain, one of which was sampled twice in autumn and spring After the insulation of the fungal strain, a total of 2921 fungal strains were identified

as the 10 dominant species with an insulation frequency of > 1.5 percent And research has shown that the colonization capacity and diversity of fungal species were

significantly higher in the spring, except for C Quercina, Acremonium sclerotigenum (F & V Moreau ex Valenta) Gam and that's D mutila species

The above theories and previous research discussed the impact of climate factors on the occurrence and diversity of fungal endophytes However, to my knowledge, how the colonization capacity of fungal endophyte will reflect under the high-temperature condition is limited

1.7 The Role of Fungal Endophytes in Sustainable Agriculture

The fungal endophytes have the potential to be important in sustainable agriculture practice according to their ability in promoting the host plant development under the abiotic and biotic stress as well as management in environmental contamination The production of phytohormones, bioactive compounds, and many agrochemical bioactive metabolites recommend the fungal endophyte to be used as cost-effective biofertilizers

1.7.1 Role of Fungal Endophytes in Controlling Environmental Contamination

Previous research elucidated studied the role of endophytic fungi in heavy metal resistant and related to the phytoremediation mechanism which is a low cost, environmentally friendly, and effective method to remove toxicants from contaminated soils In this research, it is described that the fungal endophytes related

to hyperaccumulators plants can tolerance metal accumulated in the roots of their host plant due to the long term adaptation

Iqbal Ahmad., Mohd Ikram Ansari, Farruk Aqil, 2016 studied the role of fungal

endophytes Aspergillus niger and Penicillium sp in Cr, Ni, and Cd bioabsorption

capacity Endophytic fungi have been isolated from long-term municipal wastewater treatment soil mixed with untreated mechanical profluence By applying alkali-treated dried and powdered mycelium, Aspergillus niger, and Penicillium sp Bioabsorption of Cr, Ni, and Cd was tested for their potential And it was concluded

that “Aspergillus niger and Penicillium sp have a promising bioabsorption capacity

of Cr, Ni, and Cd from single and multi-metal solutions”

Another study examined the role of dark septate endophytic (DSE) fungi in the control of contaminated environments Ousmane Diene et al., 2014 studied the role

of DSE fungi in Cesium management following the Fukushima Daiichi Nuclear

Power Plant accident in 2011 The result showed that P ibarakiensis isolates I.4-2-1, unidentified taxon isolates 312-6, and V simplex Y34 isolates in symbiosis with

Chinese cabbage seedlings showed improved plant biomass of 49 percent, 64 percent, and 82% respectively under Cs of 5ppm, but there was no increase in biomass below 10ppm Symbiosis with tomato seedlings showed an increase in biomass of 96 percent and 122% under Cs [5ppm] Under Cs [10ppm], a symbiotic tomato plant And it was concluded that the selected DSE fungi are likely to function in radionuclide-polluted environments by regulating the bioactivities of Cs

1.7.2 The Role of Fungal Endophytes in Biotic and Abiotic stress resistance

Biotic and abiotic factors determine the plant development, and yield (Bohnert et al 1995; Hamdia & Saddad 2010; Atkinson & Urwin 2012) Plant’s interactions with other organisms, which can be beneficial or harmful can be defined as biotic factors

Some symbiotic relationships with fungi are benefited to the host plant that they provide nutritional support or contribute to defense against damage from herbivores, but other symbioses with fungi are pathogens or parasites (Atkinson and Urwin 2012) Environmental factors such as temperature, humidity, light intensity, as well

as water, mineral, and CO2 availability are the abiotic stress to plants These abiotic factors influence plant growth parameters and resources Plants need an optimum level of biotic and abiotic factors to achieve maximum growth and yield

Abiotic Stress: Drought stress, a combination of reduced water content and

diminished water potential, is one of the major constrain to crops growth and yield which lead to turgor reduction, increased stomatal closure, and thus reduced cell growth and development, (Jaleel et al.,2009) Water deficiency can inhibit the photosynthesis and metabolic processes including respiration, translocation, ion uptake, carbohydrate metabolism, and nutrient uptake in plants and lead to plant cell death (Jaleel et al 2009 and Farooq et al., 2009)

Mechanism of abiotic stress tolerance by fungal endophytes can be formed by three types, (Malinowaki and Belesky, 2000): 1) Accumulation and translocation of assimilates, 2) Maintenance of cell wall elasticity and 3) Osmotic adjustment Through the first mechanism of accumulation and translocation of assimilates, when the plant is under the abiotic stress, for example, changes during the time spent photosynthesis, adjustments in the carbon metabolism and level of starches (sugar) are seen, (Gonzalez et al., 2009) The evidence of this mechanism was confirmed by the process that the plants with fungal endophytes symbiosis produce a higher amount

of soluble sugars, for example, glucose and fructose in the leaf blades, (Richardson

et al., 1992) The fungal endophytes may provoke the host plant during the metabolic process to secrete soluble sugars through which the plant gives out wall elasticity and osmotic adjustments during the condition

Rangga Yuspradana et al, 2017, studied the role of endophytic fungi in promoting rice growth under drought stress conditions They proved that endophytic fungi for

drought-resistant in rice plant by testing Acremonium sp., Penicillium sp., Nigrospora

sp., and Curvularia sp by checking the leaf color, germination capability, and

colonization capability under the three different water availability conditions (wet, moderate, and dry conditions by giving water 100, 50 and 25% of field capacity, respectively) and the result proved that these selected species of endophytic fungi are drought resistant according to the three parameters

Kumkum Azad, 2016, tested the role of fungal endophyte in drought and salt conditions-resistant by applying the symbiosis between tomato (Solanum Lycopersicum var Rutgers) and Saskatchewan saline endophyte strains For the endophyte isolation, samples were collected during summer, and for the salt-resistant test, NaCl and Hoagland’s solution was used and applied to two-week-old plants for

20 days And for the drought stress test, plants were rehydrated for 2 days after each round of drought stress As a result, following the NaCl test, the plant with endophyte colonization tended to result in a 30-50 % higher shoot biomass than non-colonized plants For the drought stress test, treatment with SK isolates Hz613, 419, and 414, Hz613 and 414 had significantly greater fresh root biomass than non-symbiosis plants during the drought test And it was concluded that treatment with Saskatchewan saline endophyte strains was able to tolerate salt or drought stress than the control treatments

Biotic Stress: “Stress that is caused in plants due to damage instigated by other living

organisms, including fungi, bacteria, viruses, parasites, weeds, insects, and other native or cultivated plants” can be defined as biotic stress (Newton et al., 2011) Biotic stress is also a major constraint that contributes to a significant loss in crop production Plants respond with the protection system to biotic stress and the mechanism can be categorized into an innate and systemic response According to Atkinson and Urwin 2012, when infection occurred, plants produce reactive oxygen species (ROS) and oxidative bursts that control the spread of pathogens Plants increase cell lignification and block the invasion of parasites to attack pathogens Also the production of natural bioactive compounds, plant protection against biotic stress

However, the mechanism that endophytes suppress plant pathogen can be through as direct and indirect effect as well as ecological effects Endophytes produce natural bioactive compounds that have antibiotics effect through which endophytes directly

suppress pathogens Fungal endophytes promote plant pathogen resistance by systematically acquired (SAR) and systemically induced (ISR) resistance SAR proteins are designed to fight plant pathogens through Salicylic acid (SA) and Pathogenesis-related (PR) proteins

According to Gunatilaka, 2006, numerous fungal endophytes produce secondary metabolites, for example, antifungal and antibacterial metabolites compounds, which emphatically hinder the development of different microorganisms including plant pathogens It was reported that a group of biocontrol strains of fungal endophytes can create antibiotics such as terpenoids, alkaloids, aromatic compounds, and polypeptides and are reported that plant pathogens are sensitive to them Hellwig et al., 2003, also proved that Alkaloids also strongly suppressed microbes In this

research, fungal endophytes Alternaria spp produced a new alkaloid of alteration

isolate, which was elucidated for having antibacterial activity against several pathogenic gram-positive bacteria Volatile oil also is one of the antibiotic compounds produced by fungal endophytes This hypothesis was proved by Atmosukarto et al., 2005 This research elucidated that the fungal endophytes

Muscodor albus, isolated from the tropical tree species, produce many volatile

organic compounds including tetrahydrofuran, 2-methyl furan, 2-butanone, and aciphyllene which have antibiotic activities

Research on the role of DSE fungi, Veronaeopsix Simplex Y34 isolated from Yaku

Island, Japan, in suppressing the disease of Fusarium, was conducted by Rida O

Khastini et al., 2012 Veronaeopsix Simplex Y3 symbiosis with Chinese chicken

showed that the disease with Fusarium wilt decreased to 71 percent and remained good The research has defined the production of mechanical resistance created by

the V simplex Y34-Hyphae network that colonized the host root and indirectly

induced a mechanism of resistance in the plant for this disease suppression activities

1.7.3 The Role of Fungal Endophyte in Promoting the Plant Growth

The fungal endophytes confer plant development through various kinds of mechanisms The activities include:

Increment accessibility of nutrients: the fungal endophytes can fix, solubilize, assemble different kinds of nutrients such as in micro or macro, and made them accessible for the host plants And thus the application of fungal endophytes can reduce the practice of chemical fertilizers It was reported that the endophytes isolated from the soybean able to solubilize phosphate, Kuklinsky-Sobral, J et al., 2004 Some endophytes isolated from the grasses grown under nitrogen-deficient soil can fix atmospheric nitrogen into ammonia (K B Rakholiya, and M D Khunt, 2015) Production of Phyto-hormone: the endophytic microorganism can produce signal molecules called phytohormones They can produce auxins, gibberellins, principally nodule-3-acetic acid (IAA), etc., which are the plant growth promoters Auxins can against ethylene and therefore, a high level of IAA with a low level of ethylene promotes plant growth and root development (Witzel et al., 2012)

Toxic molecules degradation: endophytes can degrade toxic and recalcitrant molecules at the rhizosphere as per their genetic machinery

The gibberellins (GAs) were reported to enhance crop growth and alleviate the harmful effects of abiotic stresses (Khan et al 2011, Crozier 2000; Davies 2010; King and Evans 2003; Pharis and King 1985) Lubna Bilal et al., 2018 researched to elucidate the mechanism of fungal endophytes in promoting plant growth of

Asprgillus fumigatus TS1 and Fusarium proliferatum BRL1 Fungal endophytes were

screened for IAA production and the strains that produce IAA were selected And concluded that the selected endophytic fungi can synthesize bioactive compounds, and assume that these compounds are fundamental in promoting plant growth

1.7.4 Dark Septate Endophytic Fungi (DSE) and their abilities

The DSE fungi belong to the class 4 of fungal endophytes and provide the host plant

in accessing the nutrient (Mandyam & Jumpponen, 2005) The DSE fungi are characterized by melanized hyphae and which make different from other endophytic fungi because many endophytic fungi have white hyphae It was reported that the DSE fungi can be isolated from almost 600 plant species from 320 genera and 114 families of lower plant and higher plant range from cold temperate regions to warmer temperate regions as well as tropical regions (Jumpponen & Trappe, 1998) Barrow

J.R Aaltonen R.E (2001) repotted that DSE fungi were better adapted to plants than aseptate fungi under certain conditions Previous studies have reported that the DSE fungi colonize the root of the host plant through horizontally and specifically in the epidermis and cortex and become microsclerotia, the typical formation of fungal mycelium serves as a food reservoir, in the cell of the root with any symptom of disease in the plant (Hashiba & Narisawa, 2005) And the hyphae of DSE formed into diverse and complex structures And it is confirmed by Abdellatif et al 2009, that as indicated by the course of hyphal development comparative with the long hub of the root and regularity of fungal cells and this outlined the multifaceted nature of DSE morphology, facilitated by the multicellular, septate nature of these endosymbionts The DSE fungi are reported that they have the ability to the impervious to biotic and abiotic stress and promote plant development Wiwiek Harsonowati, et al, 2020

reported that the DSE Cladophialophora chaetospira SK 51 (OGR3) can mitigate the

Strawberry Fusarium wilt disease and promote plant development Nurdebyandaru et

al., 2013 reported that the DSE fungi Veronaeopis Simplex Y34 could promote the

growth of chili plants up to 35ºC in the context of drought stress through water uptake Moreover, a meta-analysis report regarding the plant response to DSE fungi reported that based on the 56 research articles during the past 40 years of 2010, none of the reports showed a negative effect of DSE fungi on the host plant Positive effect on the host plant according to the parameters of the biomass of root and shoot or total biomass, shoot nitrogen, and phosphorous contents And these parameters were increased by 26-103% relative to control treatments The total biomass increase of 58-138% has occurred in the plants in the absence of additional inorganic nitrogen

The DSE fungi Phialocephala fortinii was reported to increase biomass, shoot

phosphorous and nitrogen concentration by 44-116% relative to non-DSE fungi symbiosis treatments, K.K Newsham, 2011 Therefore, I hypothesized that the Dark Septate Endophytic (DSE) can colonize with the Japonica rice species under the high continuous temperature and promote plant growth according to their ability of biotic and abiotic stress-resistant and promote plant growth mentioned above