Effect of salicylic acid and effective microorganism on growth and yield of mung bean in autumn winter 2020

SUMMARY Objectives Research on the effects of Salicylic Acid and effective microorganisms on the growth, development and yield of mung bean in the autumn-winter 2020.. Research Methods

VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE

Supervisor : PhD PHAM TUAN ANH

Student code : 611703

Ha Noi – 2021

DECLARATION

I declare that the thesis is the result of my own research The data and results mentioned in this thesis are honest and not used in any published thesis, dissertations, and scientific research projects previously

I hereby commit that the information cited in the thesis ensuring cited as prescribed I bear full responsibility for these reassurances

Hanoi, February 2021

Student

Nguyen Thi My Linh

Very special thanks go out to all the teachers in the Faculty of Agronomy, especially the teachers in the Department of Plant Physiology create facilitate conditions and have many valuable ideas to help me in the course of graduation thesis

The last but not least, I would like to express the gratitude to my family and my colleagues for the support they provided me through my study This would not have been possible without their support and helping

Hanoi, February 2021

Student

Nguyen Thi My Linh

CONTENT

DECLARATION i

ACKNOWLEDGEMENTS ii

CONTENT iii

LIST OF ABBREVIATIONS v

LIST OF TABLES vii

LIST OF FIGURES viii

SUMMARY viii

PART I GENERAL INTRODUCTION 1

1.1 Background 1

1.2 Objectives and requirements 2

1.2.1 Objectives 2

1.2.2 Requirements 3

PART II LITERATURE REVIEW 4

2.1 Overview of mung bean 4

2.1.1 Orign 4

2.1.2 Charateristic 5

2.1.3 The roles 6

2.2 Ecological requirements of mung bean 7

2.3 Situation of mung bean research in the world and Vietnam 9

2.3.1 Situation of mung bean research in the world 9

2.3.2 Situation of mung bean research in Vietnam 11

2.4 Research about Effective Microorganisms (EM) and Salicylic acid (SA) 13

2.4.1 Research about Effective Microorganisms (EM) 13

2.4.2 Research about Salicylic acid (SA) 17

PART III MATERIALS AND METHODS 21

3.1 Materials 21

3.2 Experiment site and research time 21

3.3 Research Contents 22

3.4 Methods 22

3.4.1 Experimental design 22

3.4.2 Cultural practices 22

3.4.3 Data collection 23

3.5 Managing collected data 26

PART IV RESULTS AND DISCUSSION 27

4.1 Effect of EM and SA on the stem height 27

4.2 The effect of EM and SA on the number of leaves 30

4.3 Effect of EM and SA on number of 1st grade branches 32

4.4 Effect of EM and Acid SA on Leaf area and Leaf area index (LAI) 35

4.4.1 Effect of EM and Acid SA on Leaf area 35

4.4.2 Effect of EM and Acid SA on Leaf area index (LAI) 36

4.5 Effect of EM and SA to SPAD 37

4.6 Effect of EM and SA on nodule formation 39

4.7 Effect of EM and SA on cumulative dry matter 42

4.8 Effect of EM and SA on photosynthetic performance 43

4.9 Effect of EM and SA on the yield and yield components 45

4.9.1 Effect of EM and SA on the yield components 45

4.9.2 Effect of EM and Acid SA on the yield 47

PART V CONCLUTIONS AND RECOMENDATIONS 51

5.1 Conclusions 51

5.2 Recomendations 52

REFERENCES 53

ANALYZING DATA IRISTART 57

APPENDIX 74

IRRISTAT Agricultural statistical software

LAI Leaf area index

LSD Least significant different

No Number

SA Salicylic acid

T Treatment

LIST OF TABLES

Table 2.1: Area, production and yield of mung beans 2014-2018 10

Table 2.2 Mung bean production situation in some countries in the world in 2017-2018 10

Table 2.3 Area and yield of mung bean in Vietnam 12

Table 4.1 The effect of EM and SA on the stem height 28

Table 4.2 Effect of EM and SA on the number of leaves 31

Table 4.3 Effect of EM and SA on number of 1st grade branches 33

Table 4.4 Effect of EM and SA on Leaf area index (LAI) 36

Table 4 5 Effect of EM and SA on SPAD index 38

Table 4.6 Effect of EM and SA to nodule formation 40

Table 4.7 Effect of EM and SA on cumulative dry matter 42

Table 4.8 Effect of EM and SA on photosynthetic performance 44

Table 4.9.1 Effect of EM and SA on the yield components 45

Table 4.9.2 Effect of EM and SA on the yield 47

LIST OF FIGURES

Figure 4.1 Growing dynamic of main stem height of mung bean 29

Figure 4.2 Growing dynamic of leaf number of mung bean 32

Figure 4.3 Growing dynamic of number of 1st grade branches 34

Figure 4.4 Effect of EM and SA on the yield 49

SUMMARY Objectives

Research on the effects of Salicylic Acid and effective microorganisms on the growth, development and yield of mung bean in the autumn-winter 2020 From there, contribute to building an intensive farming process to increase mung bean yield in Vietnam

Research Methods

Effect of Salicylic and effective microorganisms on growth, development and yield of mung bean in autumn-winter 2020 were arranged in complete randomized blocks (RCBD- Randomized Coplete Bock Design) with 6 formulas, each with 3 replicates, each of the above formulas was sown 1 plot, the total number of experimental plots is 18 plots, the area of each plot is 6m2 RCBD - Randomized Coplete Bock Design with one factor is comparison between the unprocessed Salicylic Acid formula and the effective microorganism with the Salicylic treatment formula and the effective microorganism

Results and conclusions

Through the process of implementing and researching the topic:

"Effects of Salicylic Acid and efefective microorganisms on growth and yield of DVXN7 mung bean in autumn-winter 2020’’, i draw some conclusions

as follows:

1 Treatment of EM and SA affected the growth indicators of DXVN7 mung bean In which, T6 formula (using EM2 and 0.75mM SA) had the highest height, the highest number of leaves, the total nodules and the highest nodule weight at immature fruits period T4 formula (using EM2) gave the highest 1 st grade branches T5 formula (using EM1 and 0.75mM SA) had the total nodules and the highest nodule weight at flowering period

2 Treatment of EM and SA had different effects on the physiological indicators of DXVN7 mung bean T5 formulas for highest leaf area The formulas T4 (using EM2) formulas gave highest leaf area index T3 formulas highest photosynthetic efficiency T6 formulas for highest dry matter and SPAD index highest

3 Treatment of EM and SA had different effects on the yield and yield components of DXVN7 mung bean The T4 formula (using EM2) gave the highest number of seeds/fruits The T6 formula (using EM2 and 0.75mM SA) gave highest weight of 1000 seeds, highest fruits/plant, highest individual yield and the highest actual yield

PART I GENERAL INTRODUCTION

1.1 Background

Mung bean (Vigna radiate (L.) Wilczek) belongs to the legume family,

grown for a long time and originated from India and Central Asia, distributed in the tropics and tropical Asia

Mung beans have a wide adaptability, quite drought tolerance and can adapt to areas with extreme conditions

Mung beans are short-term industrial crops with high nutritional value, easy to grow and high economic efficiency for humans Nutrients source from seeds is rich in protein (23.4%), carbohydrates 53.1%), phosphorus, potassium, magnesium and vitamins B1, B2, PP, C (Institute of Nutrition, 2007) and acids Essential amines are irreplaceable for humans The legume group makes

an important contribution to the soil ecosystem by the nitrogen-fixing symbiotic bacteria in the roots From mung beans can be processed into many different delicious foods such as mung bean cake, mung bean milk ; works in medicine

to cure a number of diseases for humans and as food for animals In addition, mung beans have a sweet, mild taste, welding properties, tonic gas, heat, cool the liver, detoxify for humans Mung bean has a short reproductive time and

is one of the three bean plants with many advantages in the production system,

so it is possible to participate in crop formulas (rotation, intercropping) to contribute to enhancing the value of land use A plant capable of improving soil fertility by fixing free nitrogen through the action of symbiotic bacteria with roots forming nodules after each crop of mung beans added to the soil 60 - 80 kg N/ha So after the soil is planted, mung beans will be more spongy, reducing costs for inorganic nitrogen fertilizers, contributing to soil protection and sustainable environmental protection

With the advantage of being short-term plants, easy to grow, providing many nutrients, improving soil and also very convenient to arrange in the formula of rotation and intercropping, so green beans are grown quite popularly However, mung bean plants are only considered as sub-crops, so the planting area is not concentrated and productivity is not high

Salicylic acid (SA) is a plant hormone plays an important role in induction

of plant defense against a variety of biotic and abiotic stresses through morphological, physiological and biochemical mechanisms SA regulates processes such as seed germination, vegetative growth, photosynthesis, respiration, thermogenesis, flower formation, seed production, senescence, and a type of cell death that is not associated with the hypersensitive response (Hayat

et al., 2010; Singh et al., 2017)

Effective Microorganisms (EM) act and interact in the soil-plant environment to suppress plant pathogens and disease, to conserve energy, to solubilise soil minerals, to aid the balance and ecology of soil microbes, and to improve photosynthetic efficiency and biological nitrogen fixation EM can improve the quality and yield of plants by reducing the incidence of pests and diseases, and by protecting against weeds, thereby contributing to sustainable agriculture

Following the fact mentioned above, we do a research on: “Effect of

salicylic acid and effective microorganism on growth and yield of DXVN7 mung bean in Autumn-Winter 2020”

1.2 Objectives and requirements

1.2.1 Objectives

Evaluate the growth, development and yield of mung bean when treated

by salicylic acid and effective microorganisms in Autumn-Winter 2020

1.2.2 Requirements

Evaluate the effects of Salicylic acid and effective microorganisms on the

growth and physiological parameters of mung beans

Evaluate the effect of Salicylic acid and effective microorganisms on the yield of mung bean

PART II LITERATURE REVIEW

2.1 Overview of mung bean

2.1.1 Orign

The mung bean (Vigna radiate (L.) Wilczek) is the third most important

bean plant after soybeans and peanuts (2 types of short-term industrial crops), native to India and Central Asia, and from there spread to many other parts of Asia

Radiocarbon archaeological evidence has uncovered traces of mung bean planted in many regions of India including the eastern part of the ancient Harappan civilization area in Punjab and Haryana dating back about 4500 years, and in The southern Indian state of Karnataka is more than 4000 years old Archaeological evidence also concluded that the green bean plant was widely planted in India about 3,500-3,000 years ago (Ho Dinh Hai, 2014)

Currently, scientists have found the ancestor of the mung bean plant, the

subspecies Vigna radiata var, sublobata), which grows wild in Mongolia That

said, Mongolia is also the place where the mung bean plant has been domesticated for a long time

In Thailand, traces of the planted mung beans and identified about 2200 years ago in the Khao Sam Kaeo area in southern Thailand

In Africa, on the island of Pemba during the era of Swahili trade, in the 9th or 10th century, traces of planted mung bean plants have also been found

Mung beans have a wide adaptability, quite drought tolerance and can adapt to areas with extreme conditions In Asia, mung beans are grown in many countries such as India, Pakistan, Bangladesh, Sri Lanka, Nepal, China, Myanmar, Thailand, Vietnam, Cambodia, Laos, Planted in Central Africa, the dry and hot regions of Southern Europe, Northeastern Australia, South America

and the Southern United States

In Vietnam mung beans are grown throughout the country from north to south This is an important vegetable and food species and a bean of special value in Vietnamese culinary culture

2.1.2 Charateristic

The mung bean (Vigna radiata (L.) Wilczek) is a legume cultivated for its edible seeds and sprouts across Asia There are 3 subgroups of Vigna radiata: one is cultivated (Vigna radiata subsp radiata), and two are wild (Vigna radiata subsp sublobata and Vigna radiata subsp glabra) The mung bean plant is an

annual, erect or semi-erect (FAO, 2012; Lambrides et al., 2006; Mogotsi, 2006)

It is slightly hairy with a well-developed root system Wild types tend to be prostrate while cultivated types are more erect (Lambrides et al., 2006)

- Stems: stem much branched, with a tendency to twine at the tips, angular, covered with long spreading hairs, reaching a height of 0.15-1.25 m and

it depends on the breed and cultivation (Mogotsi, 2006)

- Roots: root system consisting of a welldeveloped taproot with deeply placed lateral roots Airy loose soil the roots can grow up to 40 cm deep, thereby the plant can drought tolerant better Mung beans root can drought tolerant good but waterlogging is very poor, especially small trees (0 -25 days after sowing) From 15 days after sowing, the nodules are formed that are a very effective to the plant

- Leaves: Leaves alternate, dark green, trifoliolate with elliptical to ovate leaflets 5-18 cm long x 3-15 cm broad, base broadly cuneate or rounded, apex acuminate, glabrous or hairy on both surfaces, distinctly 3-veined from the base, the lateral leaflets unequal-sided

- Flowers: The pale yellow flowers are borne in clusters of 12–15 near the top of the plant Self-pollination occurs, so insects and winds are not required

- Fruit: mung bean fruit is straight cylindrical, hairy From bloom, the fruit begins to grow and matures after 18-20 days The fruit is green, more hairy, when the old is dark green and when ripe is black and less hairy Each fruit has about 5-15 seeds

- Seeds: The seeds are variable in colour, they are usually green, but can also be yellow, olive or brown, the purplish brown or black, mottled and/or ridged Seed colours and presence or absence of a rough layer are used to distinguish different types of mung bean The weight per 1000 seeds is 30 - 70g

The mung bean (Vigna radiata L.) is one of the most important edible

legume crops, is consumed all over the world, especially in Asian countries, and has a long history of usage as traditional medicine It has been known to be an excellent source of protein, dietary fiber, minerals, vitamins, and significant amounts of bioactive compounds, including polyphenols, polysaccharides, and peptides, therefore, becoming a popular functional food in promoting good health For those individuals who cannot afford animal proteins or those who are vegetarian, the mung bean is of a comparatively low-cost and has a good source

of protein for them Furthermore, mung bean protein is easily digestible, as compared to protein in other legumes (Yi-shen Z, Shuai S, 2018)

In addition to the nutritional properties of the mung bean, the

Compendium of Materia Medica (the “Bencao Gangmu”), a well-known

Chinese pharmacopoeia, has recorded that it can be utilized as traditional medicine for its detoxification activities, recuperation of mentality, ability to alleviate heat stroke, and regulation of a gastrointestinal upset Interestingly,

apart from the ancient description, recent studies have identified many other potential health benefits of the mung bean, such as its hypoglycemic and hypolipidemic effects and its antihypertensive, anticancer, anti-melanogenesis, hepatoprotective, and immunomodulatory properties beyond meeting basic nutrient requirements (Liyanage et al., 2018)

Mung bean is one of the important crops with the ability to improve soil fertility through N fixation by symbiotic association with rhizobia present in root nodules Therefore, the land after planting mung bean will become soft and more nutritious, increase the productivity of the later crop (Jat et al., 2012) Dry land areas are experiencing low agricultural yields due to severe water shortages and salinity, leading to food scarcity Mungbean is gaining attention as a short-season crop that can tolerate dryland conditions and suitable with many type of land The economic benefits of mung beans when rotated with some other crops such as groundnut have also been demonstrated (Arif and Malik, 2009)

2.2 Ecological requirements of mung bean

- Temperature:

Mung bean is a warm season crop requiring 90–120 days of frost-free conditions from planting to maturity (depending on the variety) The optimum temperature range for growth is between 27 °C and 30 °C (Duong Hong Dat, 2006) This means that the crop is usually grown during summer Seed can be planted when the minimum temperature is above 15 °C Mung bean is responsive

to daylight length Short days result in early flowering, while long days result in late flowering However, mung bean varieties differ in their photoperiod response Mung bean is considered to be heat and drought tolerant

-The light:

Mung bean is short-day plant and most are sensitive to conditions, time of day light smaller 12 hours The first sign of this reaction is that if the lighting time is too long will cause prolonged period of growth and flowering slows The

large day length influences the period of growth, it causes the flowering time to prolong and slows the ripening of the fruit, so on the same tree at the same time there are buds, flowers and green fruits, ripe fruit

For light intensity, mung bean is bright loving plant, number of sunshine hours must to reach 180-200 hours/month During the flowering period, the number of sunshine hours must to reach > 200 hours/month So hours of sunlight falls below 150 hours /month lead to plants will be weak, increases fall flower rate, insect variety So when arrangement to plant mung beans in systems intercropping with other crops should allocate time to do so when the mung beans have flower, fruit and body and leaves thrive, they will not be blocked the light from to the main tree's leaves The actual production was found to productivity of mung beans are usually higher than spring season & -winter-autumn season, at the southern provinces is higher than the north (Tran Dinh Long, Le Kha Tuong, 1998; Duong Hong Dat, 2006)

-Humidity and rainfall:

Mung bean has good drought tolerance and poor water resistance The soil humidity is suitable for mung bean growth about 70-80% Humidity and raining have a great influence during the period of flowering and making fruit Adequate rainfall is required from flowering to late pod fill in order to ensure good yield Late plantings which result in flowering during the high temperature-low moisture period in July and August will reduce yield High humidity and excess rainfall late in the season can result in disease problems and harvesting losses due to delayed maturity.The required amount of raining for the mung bean crop is 400-600 mm

Mung bean seeds are small, so seeds are sensitive to soil humidity during the budding period The appropriate humidity is 70-80% and the humidity uniformity determines the time, germination rate and uniformity of mung bean fields during the growing period The period when small plant has drought

tolerance that is considered the best Relative drought in this period makes condition for deep development root, increasing drought tolerance resistant for later growth stages During growth period, plants are sensitive to humidity The lack of humidity in this period increases the falling flower rate and drop fruit

-Soil and nutrition:

Mung beans is best suitable on fertile, sandy loam soils with good internal drainage and a pH in the range of 6.3 and 7.2 Mung bean requires slightly acid soil for best growth If it is grown in rotation, lime to attain pH of the most acid sensitive crop Root growth can be restricted on heavy clays Mung bean doesn’t tolerate saline soils and can show severe iron chlorosis symptoms and certain micronutrient deficiencies on more alkaline soils PH <5 will reduce the formation of effective nodules (Tran Dinh Long, Le Kha Tuong, 1998; Duong Hong Dat, 2006)

Mungbean has phosphorus, potassium, calcium, magnesium and sulfur requirements similar to other legumes which must be met by fertilizer additions

if the soil is deficient in these elements Despite being legumes, mung bean still need to be added with a certain amount of protein, especially for bad areas, because of the protein due to bacterial of nodules level is not sufficient enough for the plant, and in the early stages when the plant has not formed nodules yet

2.3 Situation of mung bean research in the world and Vietnam

2.3.1 Situation of mung bean research in the world

Due to the increasing demand for human nutrition, protein rich products such as mung beans are very interested and expected Therefore, the area planted

to mung beans around the world has been expanded and increased significantly

As shown in Table 2.1 below, it is estimated that the green bean area from 2014

to 2016 increased to about 67,000 ha

Table 2.1: Area, production and yield of mung beans 2014-2018

(Source: FAOSTAT,2019)

Mung bean yield also increased significantly, from 2014 to 2018 increased by about 1.3 tons / ha, the highest was in 2018 with 15.79 tons / ha Because the annual area increased, the production also increased, specifically, in

2014, it was only 21.7 million tons, but in 2018, it increased to 24.75 million tons; an increase of more than 3.05 million tons compared to the previous years

Based on the area and yield of mung beans increased over the years, scientists have researched to create new resilient, high-yielding varieties to apply to different regions of the world Some regions have applied scientific and technical advances to cultivating and ttested varieties and yielded quite high, below is table 2.2 showing acreage, production and yield in some countries in the world:

Table 2.2 Mung bean production situation in some countries in the world

in 2017-2018

National

Area (million ha) Production (tons / ha) Yield (million tons)

2017 2018 2017 2018 2017 2018 China 0,67 0,68 28,7 29,2 19,3 19,9 India 0,24 0,25 2,82 2,83 0,06 0,07 Indonesia 0,123 0,122 7,53 7,65 0,093 0,094 Thailand 0,16 0,16 1,86 1,87 0,03 0,03 America 0,09 0,09 3,1 3,2 0,003 0,003

As of 2018, China is a country with a large mung bean cultivation area and productivity and yield are quite high, from 2017 to 2018 area increased by 0.01 million hectares but the yield increased to 0.5 tons / year We can be seen that China has applied very well scientific and technical advances and is interested in breeding research to improve productivity on a large scale Other countries such as India, Thailand and the US all have small production areas, leading to low productivity and yield, and many differences compared to China

So recently, many countries around us such as India, Thailand, the Philippines, etc have paid attention to creating green bean varieties with yields

of 10-12 quintals / ha or more, large seeds, color The seeds are beautiful, have a short growth time, are ripe relatively concentrated, and have quite resistance to major pests and diseases

2.3.2 Situation of mung bean research in Vietnam

In Vietnam, mung bean has been grown for a long time, throughout the country, one of the traditional crops with many purposes: getting seeds, improving soil, preventing erosion, making green manure But now, cultivated area is limited, scattered from South to North, from delta provinces to midland and mountainous areas Mungbeans are not considered as the main crops, only intercropping, crop planting, to take advantage of land, increase income, so the area and productivity of mung beans in Vietnam is not high

The area of mung bean production in Vietnam for 4 years of 2012 and 2015 varied from 88.180 – 98.200 ha, the area of mung bean production in 2015 decreased compared to 2012 was 7250 ha The average mungbean productivity of Vietnam over 4 years varies from 1026 - 1098 kg/ha The average mung bean productivity of the whole country tended to increase gradually over the years and the highest yield in 2015 was 1098 kg/ha

Mung bean production area in Vietnam is developed in 7 ecological regions across the country (Table 2.3) The area of mung bean production between regions

in 2015 varied from 4880 - 25120 ha, of which 3 regions with large mung bean production areas are the Central Highlands, North Central and South Central Coast respectively: 25120 ha; 18470 ha; 18090 ha

Table 2.3 Area and yield of mung bean in Vietnam

Region

2012 2013 2014 2015 2012 2013 2014 2015 Red river delta 4.10 4.80 4.51 4.88 1463 1458 1452 1511

Mekong Delta 8.30 6.60 7.28 7.76 1506 1576 1591 1719

Country 98.20 93.80 88.18 90.95 7981 7997 8235 8451

(Source: National Institute of Agricultural Planning and Projection, 2016)

The average mung bean yield in the ecological regions has a big difference Mung bean yield fluctuated among regions in 2012 from 837 - 1506 kg/ha; 2015 varied from 861 - 1719 kg/ha Average of mung bean yield reached the highest in the Mekong River Delta and Red River Delta 1719 kg/ha and 1511kg/ha (2015)

The average mung bean yeild reached the lowest in the Central Highlands and the North Central In 2015, the mung bean yeild in the North Central was 938 kg/ ha, lower than the national average yield of 160 kg / ha The Central Highlands

mung bean productivity reached 861 kg/ha and lower than the national average productivity of 236 kg/ha Mung bean yield of the country in general and the North Central region in particular over the years tends to increase, which is the application of new mung bean varieties and new farming techniques to production

Production of beans in general and mung bean in particular is increasingly playing an important role in the process of restructuring plant structure and promoting commodity production in our country Compared to some other crops in the same conditions, mung bean is more effective, easier to consume Therefore, it

is necessary to pay attention to the development of mung bean to expand the area, contributing to increasing production and promoting the development of the country's economy

2.4 Research about Effective Microorganisms (EM) and Salicylic acid (SA)

2.4.1 Research about Effective Microorganisms (EM)

 Research application of Effective Microorganisms (EM) in the world

EM consists of 80 species of rare and anaerobic organisms, selected from more than 2000 species commonly used in food technology and fermentation technology (Pham Thi Kim Hoan, 2008) EM products were born and quickly absorbed and applied in many fields by countries around the world Efective Microorganisms Research Organization (EMRO) is established in many countries around the world and has close relationship with EMRO in Japan (Pham Van Ty, Vu Nguyen Thanh, 2006)

Through scientific reports at international conferences on technology, it is shown that EM can increase the biosphere balance, increase the diversity of agricultural land, enrich the soil components to improve crop quality Therefore,

EM is welcomed by countries around the world as a solution to ensure sustainable agricultural development while protecting the environment

Thailand hosted the First International Conference on Redemptive Nature Agriculture and EM agriculture in October 1989 Scientists discussed the value

of EM technology and encouraged its use Thanks to that, the Asia Pacific Natural Agriculture Network (APNAN) was established, a non-governmental organization that aims to promote research, development and practical application of solutions technology with natural agriculture associated with technology and Effective Microorganisms EM (Pham Thi Kim Hoan, 2008) The 2nd International Conference held in Brazil in October 1991 had a series of reports on the effectiveness of EM on the growth, development, yield and quality of some crops such as: rice, potatoes, sweet potatoes, in Brazil, Japan, Korea, (Pham Thi Kim Hoan, 2008)

New research on EM and EM applications around the world has been published at conferences such as studying the effect of EM on seed germination, the effect of EM on the growth, development and yield of some crops such as rice, maize, soybean, etc In addition to the positive results on crops, there are also promising studies on the effectiveness of EM in livestock, aquaculture, waste treatment, Dr James F.Par Department Agricultural Research - The US

Department of Agriculture said: “We recognize EM technology is a potentially

valuable tool that can help farmers develop economically, environmentally and socially sustainable farming systems ”(Higa, Parr, 1994)

With effective research results, more than 150 countries have implemented

EM technology and it is being produced by more than 50 countries Many EM factories and factories are built in many countries and regions, each year thousands of tons of EM are produced such as: USA, Thailand (1000 tons / year), Brazil, Japan (1200 tons / year), (Pham Thi Kim Hoan, 2008)

 Research and application of Effective Microorganisms (EM) in Vietnam The research works on Effective Microorganisms are conducted from the early years of the 1960s in Vietnam, it was not until after the 1980s that it was officially included in the state-level scientific programs such as: "Biology for

Agriculture" period 1982-1990; program "Biotechnology" KC.08 period 1991 - 1995;

In 1997, research institutions and some localities such as Hanoi University

of Agriculture, Hanoi National University, Plant Protection Institute, Thai Binh Province, conducted initial tests EM preparations in the fields of cultivation, plant protection, environmental sanitation, and seeing certain positive effects of

EM technology In 1998, the Ministry of Science, Technology and Environment decided to carry out an independent state-level project: “Research, experiment and absorb EM technology in the fields of agriculture and protection environmental biology since 1998 2000 led by Prof Dr Nguyen Quang Thach The thesis assesses the safety of EM preparations, determines the variable ingredients and characteristics of EM preparations, the effectiveness of EM preparations in waste treatment, environmental sanitation, cultivation and breed Since then, there have been many researches on applying EM technology in institutes, centers, provinces and cities, especially in the field of environment The research results on the application of EM inoculants on some Vietnamese crops confirmed that EM increases productivity and quality, and at the same time limits the increase of some diseases such as blight and blight in rice plants EM has the effect of shortening the growing time, increasing yield and quality of rice For soybean plants, EM treatment increases germination rate, stimulates root development, increases chlorophyll content, increases tolerance

to adverse external conditions (drought, salinity, waterlogging, )

At Vietnam National University of Agricultural, EM treatment for rice yield increased by 8-15% and no sheath blight In soybean, using EM in spray or soil application mode increases seed germination rate, increases chlorophyll content and reduces root rot

- Research results of Efective Microorganisms

Based on the operation and blending principle of Efective Microorganisms (EM), the Institute of Agricultural Biology - Vietnam National University of Agriculture has produced EM The product has been tested and applied effectively in many fields and environments The inoculant has the same quality

as imported EM, but has reduced production cost by 1/3, so it has been widely used in the domestic market with the consumption of thousands of liters a year (Pham Thi Kim Hoan , 2008)

- Characteristics of EM:

The strains of microorganisms in the E.M inoculant were isolated in very harsh environmental conditions such as: temperature, high pressure, low pH After that, they were fed again and tempered in harsh environmental conditions Therefore, the microorganism strains contained in the E.M inoculant have strong vitality, have a very high tolerance to adverse environmental conditions, have very high activity and efficiency

- Function of EM:

+ EM effects on soils and crops

EM has been used on many different soils and crops over a wide range ofconditions Results show that in most cases EM gives positive results EM is not a substitute for other management practices EM technology is an added dimension for optimising our best soil and crop management practices such as oncrop rotations, use of composts, crop residue recycling, and biological control

of pests It used properly EM enhances soil fertility and promotes growth, flowering, fruit development and ripening in crops It can increase crop yields and improve crop quality as well asaccelerating the breakdown of organic matter from crop residues The population of beneficial micro-organisms in the soil is also increased helping to control soil diseases through competitive exclusion.In New Zealand EM has Bio-Gro certification as an “Approved organic product”

+ EM for weeds pests and diseases

EM is not a pesticide and contains no inorganic chemicals EM is a microbialinoculant that works as a bio-control measure in suppressing and/or controllingpests through the introduction of beneficialmicroorganisms to soils and plants Pests and pathogens are suppressed or controlled through natural processes byenhancing the competitive and antagonistic activities of the microorganismsin the EMinoculants

2.4.2 Research about Salicylic acid (SA)

Salicylic acid is a simple phenolic compound synthesized in a wide range

of prokaryotic and eukaryotic organisms, including plants (Raskin, 1992) Leaf

and bark of willow tree (Salix sp.) contain large amounts of SA, which was

widely used as a medication for pain relief in the ancient world In 1828, German scientist Johann A Buchner purified salicyl alcohol glucoside (SA derivative called salicin) from willow bark Ten years later, an Italian chemist Raffaele Piria working in Paris converted salicin into an acidic aromatic compound that he named salicylic acid In 1859 Hermann Kolbe et al chemically synthesized SA, but the bitter taste and side effects limited the long-term use of SA as a medication

In 1979 a role of SA in the defense response of plants was discovered Treatment of tobacco plants with Aspirin resulted in enhanced resistance to tobacco mosaic virus (White, 1979) Subsequent studies revealed that pathogen infection increases the level of SA and promotes transcription of genes encoding pathogenesis-related proteins in plants, which confers disease resistance

In addition to defense responses, SA is implicated in the regulation of a variety of biological processes, such as seed germination, seedling development, nodulation in legumes, plant vegetative growth, senescence-associated gene expression, flowering time, fruit yield, respiration, as well as response to ultraviolet (UV)-B radiation, ozone, metals, drought, temperature, and salinity stresses (Khan et al., 2015; Vlot et al., 2009)

Salicylic acid is often found in vascular plants, playing an essential role in regulating plant growth and responding to stress SA affects the physiological and biochemical activities of plants and can play an important role in regulating growth and productivity In addition, SA also plays a role in establishing protective responses against various bacterial infections and resistance in plants Able to transport from healthy tissue and organs but SA is not a signal of disease transmission For flowering, when treated with SA, it produces more early flowering and flowers than not treated Some reports indicate that SA increases leaf area and dry weight in maize and soybean For wheat before sowing, treating SA before sowing increases germination rate, promoting growth SA reduces Cd toxicity in maize and barley AS reduces prolin accumulation in wheat to increase tolerance to salinity Exogenous SA treatment significantly improved salinity tolerance in corn However, when used in higher concentrations have inhibitory effects

According to Nguyen Thi Phuong Dung et al., (2016), SA treated exogenously or highly synthesized in tissues also helps plants resist abiotic stresses such as hot, cold, and salt The impact of SA on resistant of crop:

- Exogenous application of SA enhances growth, affects flowering and crop yields

- The exogenous application of SA causes the immune system in plants, thereby providing a significant protection against various stresses

- SA is effective at reducing the toxic effects arising in plants due to exposure to many abiotic stresses such as heavy metals, temperature, water, ozone, UV and saline radiation…

- Exogenous application of SA enhances photosynthesis and many other biochemical physiological characteristics of plants However, higher concentration can cause stress in plants

- Exogenous application of SA enhances the operation of the system antioxidant enzyme

- SA activates protective genes when plants are attacked by pests and produces protective substances such as PR1, PR2 and PR5 SA also plays a very early role when plants are attacked by pests, creating a defense mechanism that immediately kills cells in the field attacked by pests

The effect of SA on exogenous treatment on Rhizobium bacteria with symbiotic nodule of legumes is considered SA affects the early stage of Rhizobium and also delays nodulation, thereby reducing the number of nodules

on the plant Nitrogen metabolism is an important aspect of legumes with SA concentrations higher than 10-3 or 10-4 inhibiting the nitrogen metabolism enzyme

It is clear that exogenous applications of salicylic acid can bring significant benefits in agricultural production and can be applied to enhance crop production

Currently, in Vietnam have been many studies in the application of SA with plants with certain results Research results of SA's effect on drought-tolerant cucumber (Nguyen Thi Phuong Dung et al., 2016) showed that adding

SA to drought formulas reduced the impact of drought on the periods of cucumber, increasing the tree height and leaf area The 2 of SA concentrations,

SA 0.25 mM is more effective than 0.5 mM Another study on SA with mungbean showed that SA treatment increases the germination rate of seeds, increases germination length, prolin content of stem, and SA treatment also reduces the level of damage caused by Al According to Phung Chi Son, SA solution at a concentration of 1000 ppm can increase resistance to anthracnose

on dragon fruit when combined with pesticides In addition, according to some studies, preparations containing SA active ingredients have ability to repel aphids, SA is also applied in the prevention of rice sheath blight

PART III MATERIALS AND METHODS 3.1 Materials

Variety: DXVN7 mung bean

- The origin:

DXVN7 mung bean was derived from crossing DX102/Vinh Bao 4 in Spring 2007 at the Maize Research Institute In recent years mung bean DXVN7

is being grown widely in many places because it has ability for high yield,

drought resistance and identified lodging resistant ability and slightly sensitive with brown spot pathogen

Growth characteristics: DXVN7 is a variety with short growth duration 65-68 days in Summer season, main stem 40 - 65 cm The yield is in range of 10

to 18 quintals/ha and it depends on intensive conditions The weight of 1000 seeds is 51 - 55 g, moldy green seeds and it is suitable for consumers' tastes The ripening of DXVN7 mung bean is focuses so convenient for harvesting, harvest 2-3 times/crop

- Fertilizer: Ure (N: 46%), super phosphorous (P2O5: 16%), kali clorua (K2O: 60%)

- Effective microorganism: microorganisms stimulate plant growth, this microorganisms is provided by the Microbial technology, Department of Biotechnology, Vietnam National University of Agriculture

- Salicylic acid

- Lime eggshell

3.2 Experiment site and research time

- Experiment site: Faculty of Agronomy in Vietnam National University

of Agriculture

- Research time: the field experiment was conducted from 9/2020 to 12/2020

3.3 Research Contents

Studying the effect of EM and SA on the growth, development and yield

of mung bean DXVN7 in the Autumn-Winter season 2020, experiment is performed in Gia Lam - Ha Noi

3.4 Methods

3.4.1 Experimental design

The experiment was laid out in a randomized compete block design (RCBD) with three time repeat, land area of each plot was 6 m2

- Number of treatments: 6 formulas

+ Treatment 1: control (no use SA and EM)

+ Treatment 2: use EM 1

+ Treatment 3: use EM 2

+ Treatment 4: use 0.75mM SA

+ Treatment 5: use EM 1and 0.75mM SA

+ Treatment 6: use EM 2 and 0.75mM SA

- Number of plots: 18 experimental plots

- Density: 35 plants/m2

- Distance: plant by plant 5 cm, row by row 35 cm

Experimental layout design

Replication 1 T1 T3 T2 T5 T4 T6 Replication 2 T3 T2 T1 T6 T5 T4 Replication 3 T2 T4 T5 T3 T6 T1

3.4.2 Cultural practices

- Soil Preparation:

Before transplanting, soil will be prepared and cleaning up, weeding, mix well

to ensure it has to be loose and uniform

- Sowing seeds and seed emergence:

Sowing the mung bean 2 - 3 cm deep into the soil, and space the seed about 15-20 cm, row by row is about 35-40 cm, For each hole put 2 seeds, after that keeping 1 tree per hole, remove weaker plants to let the stronger plants flourish, ensuring the density 15-20 trees/1m2

- Fertilizer application:

Basal application: 100% P2O5 + lime eggshell (200 kg/ha)

Top dress application 1st: 50% K2O + 50% N after plant have 2-3 real leaf Combine weeding, cultivating soil

Top dress application 2nd: remaining amount of fertilizer apply after plant have 4-5 real leaf Combine weeding, tillage, digging to prevent from falling Spray leaves through 3 stages:

+ 1st time: when the plant has 3-4 real leaves (300 liter/ha)

+ 2nd time: when the plant flowering 50% (450 liter/ha)

+ 3rd time: the plant produces young fruit, after 10 day stop flowering (600 liter/ha)

- Watering: ensuring plants are watered enough and frequently Keep land

regularly about 70-80 % field moisture content, limit watering during the period

of seed maturity and fruit maturity

- Weed and disease control: During experiment, observing plants

frequently and giving out prevention and treatment in time

- Harvest: harvest in accordance with the maturity of the fruit, drying and

preserving the seeds During storage, attention should be paid to check for pests

3.4.3 Data collection

- Indicators of growth time

+ Time from sowing to germination: time from seeds sowing to have 50%

of the plants grow out of the ground and appear 2 cotyledons

+ Time from sowing to flowering: the number of days from sowing to having 50% of plants with at least 1 flower

+ Total time of growth: the number of days from sowing to end of harvesting

- Indicators of growth and development

+ Main stem height (cm): 5 plants/1 plot was measured height from cotyledons to the top of the main stem (7 days/times)

+ Number of 1st grade branches: 5 plants/1 plot was counted the number

of 1st grade branches on main stem (7 days/times)

+ Number of leaves on main stem: 5 plants/1 plot was counted the number

of leaves on the main stem (7 days/times)

+ Fresh weight of whole plant (g/plant): follow 3 periods of flowering, immature fruits and at 1st harvest fruits, randomly 3 plants/replication were collected, root was cleaned then weight

+ Dry weight of whole plant (g/plant): follow 3 periods of flowering, immature fruits and 1st harvest fruits, randomly 3 plants/replication were collected, dried in oven until stable weight is obtained and weight to have dry matter data

+ Number of total nodules (nodules/plant): though 3 periods of flowering, immature fruits and 1st harvest fruits, randomly 3 plants/replication were collected then number of total nodules was counted

+ Effective nodules rate (%): total number of effective nodules (pink in colour)/total number of nodules x 100

+ Weight of total nodules/plant (g/plant): 3 plants/replication were collected then weight of total nodules per plant

- Indicators of physiological

+ Leaf area index (LAI): Follow 3 periods of flowering, immature fruits and 1st harvest fruits, each plot taked random 3 plants, identified the leaf area by

weighing method Weight all the fresh leaves of the plant gave P1 (g) 1dm2 of fresh leaf was cut then weight gave P2 (g) Leaf area equal P1/P2 (dm2/plant) then determined leaf area index (LAI)

+ SPAD value: SPAD 502 indicator gauge was used to measure SPAD index After 7 days sprayed EM and SA, 5 plant/replication were selected and measured the third leaf from the top down

+ Photosynthetic performance: the amount of dry matter accumulated over 1 m2 during 1 day and night This indicator relates to the cumulative capacity of the crop so it reflects the productivity of the plant correctly

Photosynthetic efficiency is calculated by the formula:

P1, P2: weight of dry matter was measured first and the second time after

T days (g)

L1, L2: leaf area was measured first and the second time after T days (m2)

- Indicators of productivity and quality

+ Number of fruits/plant: total fruits of 10 plants/1 plot were counted and calculated the average fruits per plant

+ Number of firm fruits/plant: total firm fruits of 10 plants/1 plot were counted and calculated the average firm fruits/plant

+ Number of seeds/fruit: counted seeds on each fruit and took the average seeds/ fruit of each treatment

+ Weight of 1000 seeds (g): 1000 seeds for 3 replicates of treatments were weighed and took the average

+ Individual yield (g/plant): individual plant was harvested and weighed seeds/plant

+ Theoretical yield (quintal/ha): individual yield x density (plant/ha)

+ Actual yield (quintal/ha): all plants in the experimental plot were harvested, seeds were cleaned from pod and weighed seeds/plot then convertion

to quintal/ha

3.5 Managing collected data

Variance ANOVA, Coefficient of Variation (CV %) and Least Significant Difference (LSD5%) for field experiments were calculated with IRRISTAT version 5.0 software All other statistical calculations were carried out with the Excel Microsoft Office

PART IV RESULTS AND DISCUSSION

4.1 Effect of EM and SA on the stem height

A stem is one of two main structural axes of a vascular plant, the other being the root The primary functions of the stem are to support the leaves; to conduct water and minerals to the leaves, where they can be converted into usable products by photosynthesis; to transport these products from the leaves to other parts of the plant, including the roots The stem conducts water and nutrient minerals from their site of absorption in the roots to the leaves In addition, stem also has functions as a support for the entire upper parts of the plant, and it is a determining factor for resistance to fall and height of plant

Stem height is an important indicator that reflects the growth and development of plants under research and production conditions It is not only influenced by the genetic nature of the breed but also influenced by external factors such as humidity, temperature, light density…etc, density, cultivation methods, pests and diseases and amount of nutrients provided to the plant

Stem height is an important biological indicator because of it affect other biological indicators such as the number of leaves, the number of branches the same breed but different care regime, the height of the plant is also different

Therefore, the study of stem height is the foundation factor for identifying appropriate cultivation techniques, creating a balance for height of mung bean,

so bring economic significance and high scientific The results height growth of mung bean were showed in table 4.1

Table 4.1 The effect of EM and SA on the stem height

Unit: cm/plant

Treatment The height of plant

22DAS 29DAS 36DAS 43DAS 50DAS T1 13.04 23.89 36.03 43.92 50.41 T2 12.85 24.24 36.05 42.9 51.09 T3 12.43 23.75 36.67 43.86 51.63 T4 13.25 24.8 36.31 44.08 51.96 T5 12.44 24.75 36.74 43.85 51.29 T6 12.94 25.9 36.99 44.11 52.41

DAS: day after sowing

Table 4.1 shows that the use of EM and SA had an impact on the growth dynamic main stem height of DXVN7 mung bean After mung bean was treated

by EM and SA, the stem height was changed:

In the early period (from sowing to 22 days) of plant without treatment, the height of plant did not has much difference Plant height ranged from 12.43 cm – 13.25 cm The formulas T4 had the highest with 13.25 cm The formulas T3 gave the lowest plant height with 12.43 cm The formulas T1, T2, T5, T6 has height of plant respectively are: 13.04 cm; 12.85 cm; 12.44 cm and 12.94 cm

At time from 29 days after sowing (DAS) to 43 days after sowing is the period of strong growth During this period, EM and SA was used so plant height increased gradually though tracking times Besides the trees blossomed and developing leaf stems, mung bean plant adapt to the environmental conditions, the roots had use avaiable nutrients and fertillzers So we can see the impact of EM and SA on plant made the plant increase height significantly In

formulas T3 had the highest height, increased 20.11 cm from 23.75 cm to 43.86

cm The formulas T1 increased at least 15.03 cm from 28.89 cm to 43.92 cm The formulas T2, T4, T5, T6 also increased respectively are 18.66 cm; 19.28 cm; 19,10 cm and 19.10 cm

From 43 days after sowing to 50 days after sowing the growth dynamic of height was stability, this is the time seeds began form In this period, plant height ranged from 50.41 cm – 52.41 cm Therein, T6 gave the heighest plant height with 52.41 cm and T1 had the lowest with 50.41 cm The formula T2, T3, T4, T5, T6 were taller than T1 sequence 0.68 cm, 1.22 cm, 1.55 cm, 0.88

cm, 2 cm but this difference was not significant

Figure 4.1 Growing dynamic of main stem height of mung bean

Thus, the following process can conclusions that: on the same ground, climatic conditions, the same conditions of care, use EM and SA affected to the height of the main stem