Nghiên cứu một số đặc điểm sinh học, sinh thái và biện pháp bảo tồn loài pơ mu (fokienia hodginsii (dunn) a henry h h thomas), sa mu dầu (cunninghamia konishii hayata) ở khu dự trữ sinh quyển miền tây

Thomas, Cunninghamia konishii Hayata IN WESTERN NGHE AN... The necessity of the thesis Western Nghe An Biosphere Reserve BR located in Nghe An province is thelargest biosphere reserve in

VINH UNIVERSITY

NGUYEN THI THANH NGA

STUDY ON BIOLOGICAL AND ECOLOGICAL

CHARACTERISTIC, AND CONSERVATION MEASURES

OF Fokienia hodginsii (Dunn) A Henry & H H Thomas, Cunninghamia konishii Hayata IN WESTERN NGHE AN

1 The necessity of the thesis

Western Nghe An Biosphere Reserve (BR) located in Nghe An province is thelargest biosphere reserve in Southeast Asia with an area of 1,299,795 hectares,comprising 9 districts, a green corridor connecting three core zones: Pu Mat NationalPark (NP), Pu Huong Nature Reserve (NR) and Pu Hoat NR This is an area withvery diverse habitats and great biodiversity value

Fokienia hodginsii (Dunn) A Henry et H H Thomas and Cunninghamia konishii Hayata are two of the 12 Pinophyta species recorded in Western Nghe An

BR They are a lot of value for human life, besides they play an important role inkeeping the watershed forest ecosystem These are two species listed in IUCN,Vietnam Red Book and Group IIA of Government Decree 32/2006/NĐ-CP

Until now, there have been a number of literary published about distribution,

biological and ecological characteristics and the natural regeneration of F hodginsii and C konishii in some region Western Nghe An BR However, there has not been

any research on the scientific basis to preserve these two conifers for the whole area

Because of the above reasons, we have chosen the topic of research: “Studying

on biological and ecological characteristics, and conservation measures of

Fokienia hodginsii (Dunn) A Henry & H H Thomas, Cunninghamia konishii

Hayata in Western Nghe An Biosphere Reserve” to contribute the preservation and

development of two rare species in Nghe An province and Vietnam

2 Research objectives

2.1 Generalobjectives

Adding more information about the biology, ecology, propagation, and

chemical composition oil of F hodginsii and C konishii in Western Nghe An BR.

This is the scientific basis that proposes solutions to preserve and develop theseconifers in the study area

2.2 Specific objectives

- Describing morphology, anatomy, characteristics of seasonal development,ecological characteristics (distribution, density, area, reserves, some biological biome

characteristics, climatic conditions, and soil) of F hodginsii and C konishii.

- Describing characteristics of natural regeneration and assessing propagative

ability by seed, cuttings of F hodginsii and C konishii.

- Identifying the chemical composition of essential oil of parts of F hodginsii and C konishii.

- Assessing the current situation and effect on F hodginsii and C konishii to

propose solutions that preserve and develop these two species

3 Scientific and practical significance

3.1 Scientific significance

Add more data on the biology, ecology, propagation and chemical composition

of essential oil of F hodginsii and C konishii The two conifers have an important

role in forest ecosystems as well as human life

3.2 Practical significance

- Providing scientific data on of F hodginsii and C konishiifor of the whole

Western Nghe An BR to orient for the conservation and development of these twospecies in Nghe An province as well as in some other areas in Vietnam

- The thesis is a reference, researching and teaching documentary for scientists,technical staff, students, about the biological and ecological characteristics,

chemical composition in essential oil and propagation of F hodginsii and C konishii.

4 New contributions to the thesis

- Adding some new data on the anatomical structure of F hodginsii and C konishii.

- Adding some new data on distribution, area and reserves of F hodginsii and

C konishii for the whole Western Nghe An Biosphere Reserve.

- Adding some new data on the use of growth regulators in propagation by

cuttings of C konishii in Vietnam.

- Adding some new data on the chemical composition of essential leaf oil of F hodginsii in Vietnam, essential cone oil of F hodginsii and essential cone oil, essential roots oil and resins of C konishii for science.

- Adding some new data on red and white timber type of F hodginsii and C konishii by some genetic parameters.

5 Structure of the thesis

The thesis includes 137 pages: Introduction (4 pages), Chapter 1.DocumentationOverview(26 pages), Chapter 2 Objects, content and researchmethodology(17 pages), Chapter 3 Results and discussion (88 pages), Conclusionsand recommendations (2 pages), The author’published list related to thesis,References, 1, 2, 3, 4, 5, 6, 7 appendices

Chapter 1 DOCUMENT OVERVIEW

1.1 OVERVIEW of Pinophyta

1.1.1 In the world

In the world, the Pinophyta currently exist in the vegetation on the earth's surface

is not much There are 615 species belonging to 70 genera, 8 families (Farjon A., 2010)

According to the Red List of the International Union for the Conservation ofNature has 211 species of conifers (34%) are estimated to be extinct at theinternational level (IUCN, 2013) The conifers in the tropical are about 200 and themost threatened habitats are tropical rainforests (Filer D and Farjon A., 2013)

F hodginsii and C konishii in the world.

F hodginsii, there have been some studies on this species that focus on

classification, morphology, anatomy and development of reproductive organs,fertilization, some ecological characteristics, propagation testing by cuttings, analysis

of leaf oil chemical composition and genetic characteristics of the diploidchromosomes of this species (Zheng Rong et al., 2007), (Li H L., Keng H., 1994),(Chen Z K and Wang F H., 1980a, 1980b, 1981), (Farjon A.,2010), (Pan J G et al.1991), (Zonneveld B J M., 2012),

C konishii is less published than F hodginsii These studies mainly focus on

classification, morphological descriptions, ecological characteristics, essential oilcomponents analysis of stem wood and leaves, propagation testing by cuttings andgenetic characteristics of the diploid chromosomes of this species (Schlarbaum S E.and Tsuchiya T.,1984), (Jia G et al.,1998), (Atkinson B A et al., 2015), (Su Y C etal., 2012), (Liang Wen-ying, 2010),

1.2.2 In Vietnam

The short history was published about the morphological characteristics,ecological and reproductive characteristics, propagation, analysing the chemical

composition of essential oils and studies on genetic characteristics of F hodginsii and

C konishii in Vietnam.

So far, in Vietnam there have been a few studies on F hodginsii studied These

studies mainly focus on distribution and general ecological characteristics, studyinggenetic characteristics of some populations in some localities, essential oil componentsanalysis of stem and root wood There were some propagated by sowing testing seeds,cuttings and tissue culture results with satisfactory result (Nghia Nguyen Hoang vàTien Tran Van, 2002), (Hiep Nguyen Tien et al., 2004), (Anh Pham The et al., 2007),(Hung Nguyen Quang et al , 2010), (Phuong Trang Nguyen Thi, 2012),

C konishii distributed in Northern (Ha Giang and Son La provinces) and

central provinces (Thanh Hoa and Nghe An provinces) The research results on thisspecies in the country are very limited, mainly focused on describing the externalmorphology, some general ecological characteristics, genetic characteristics of somepopulations and plant propagation test by sowing seed (Phuong Trang Nguyen Thi,2012), (Loc Phan Ke et al., 2013), (Thai Tran Huy et al., 2007),

1.2.3 In Western Nghe An Biosphere Reserve

There are a few studies on F hodginsii and C konishii in the Western Nghe An

Biosphere Reserve These studies mainly stop at the discovery of a number ofdistribution points, describing common biological and ecological characteristics Therearen’t any studies on these two species in a comprehensive and systematic manner,especially on a large scale such as the whole Western Nghe An BR (Loc Phan Ke et al.,2007), (Sinh Nguyen Van, 2009), (Sam Hoang Van and Dung Tran Duc, 2013),

1.3 Natural conditions, Economic and Social characteristics of the studying area

The contents of this section include geographic location, topography, climaticcharacteristics, hydrological, soil, socio-economic characteristics of the studying area

Western Nghe An BR has a large area (1,299,795 ha), located in 9 districts:Que Phong, Quy Chau, Quy Hop, Ky Son, Tuong Duong, Con Cuong, Anh Son,Thanh Chuong, and Tan Ky It has 440,8 km of Vietnam - Laos borderline

This biosphere reserve is located in the North Truong Son range with slopingterrain, mountainous terrain, in general, decreasing from the North West to the SouthEast, the tropical monsoon climate, there are many major soil groups such as FerralicAcrisols, Humic Acrisols, Fluvisols, …

The Biosphere Reserve is inhabited by a number of ethnic groups, with apopulation of 1,197,628 (accounting for 41% of the province's total population)(2015) The income of people in the area is mainly from agricultural production, theliving standard of people in the region is very low (estimated at about 16 millionVND/person/ year)

Chapter 2 OBJECTS, CONTENT AND RESEARCH METHODOLOGY

2.1 Object and scope of research

Research object for the theme is Fokienia hodginsii (Dunn) A Henry et H H Thomas and Cunninghamia konishii Hayata, Cupressaceae, growing naturally in 6

districts: Que Phong, Quy Chau, Ky Son, Tuong Duong, Con Cuong and Anh Son inWestern Nghe An BR, Nghe An Province The research was conducted from October

2013 to June 2017

2.2 Research content

The theme focus on the following content:

- Researching on morphology, anatomy, seasonal development of F hodginsii and C konishii.

- Researching on ecological characteristics, including distribution, density,area, reserves, topographic, soil, sun drying, climate, some biome characteristics(structural level, forest cover, ecological relationship with species that grow together)

of F.hodginsii and C konishii.

- Assessment of reproducibility and testing of propagation techniques by seeds

and cutting of F hodginsii and C konishii.

- Analyzing the chemical composition of the essential oil of the plant parts

- Analyzing some genetic parameters of red and white wood of F hodginsii and

C konishii.

- Assessing the situation and proposing solutions to the conservation and

development of F hodginsii and C konishii in Western Nghe An BR.

2.3 Research methodology

2.3.1 Inherited document method

Inheriting selective data sources, the published literature with related content ofresearch thesis

2.3.2 Interview method

The interviewed method is applied according to Gary J Martin (2002)

2.3.3 Fieldwork method

Fieldwork methods are applied according to "Methods for Plant Research”

(Thin Nguyen Nghia, 2007)

2.3.3.1 Used instruments and equipment

Using survey instruments and equipment in the terrain such as GPS, camera,height meter,

2.3.3.2 Identification of survey location and routes

a Survey routes

Establishing 47 multi-terrain survey routes on mountain ridges, along with themountain ridge, along main streams, and through villages located in 22 communes of

6 districts in the Western Nghe An BR include:

- Que Phong district has 18 routes: 1 Pung stream, 2 Nam Can stream, 3.Muong Phu hamlets, 4 Lan stream, 5 Huoi Banh Thong stream, 6 Mit stream, 7.PhaPhay ravine- Nui Lua ravine, 8 Huoi Cham stream, 9 Huoi Hap stream, 10 HuoiGiai stream, 11 Huoi Dum stream, 12 Huoi Linh stream, 13 Pha Loi mountain, 14.Chop Chap mountain, 15 Pu Hoat mountain, 16 Kia ravine, 17 Huoi Que ravine 18.Huoi Huong ravine (another name is Nga Ba Pu Lon)

- Quy Chau district has 3 rotes: 1 Muc Pan hamlets, 2 Pa Ha ravine, 3 PuLon mountain (Dien Lam commune)

- Ky Son district has 9 routes: 1 Huoi Xa ravine, 2 Lot ravine, 3 Pu Lonmountain (Tay Son commune), 4 Pung hamlets, 5 Pha Noi hamlets, 6 Buoc Muhamlets, 7 Na Ca ravine, 8 Huoi Lom ravine, 9 Nam Khien ravine

- Tuong Duong district has 8 routes: 1 Loang Quang mountain range, 2 PhoBen mountain, 3 Phu Pha Den mountain range, 4 Ngan ravine, 5 Pha Lom hamlets,

6 Da ravine, 7 Thoi ravine – source of Bu ravine (include small Pu Xam Liemridge), 8 Pu Đon Can mountaintop

- Con Cuong district has 8 routes: 1 799 adjacent road, 2 Luong ravine, 3

808 Borderline, 4 Pu Nhong mountain, 5 Kem ravine, 6 Cong ravine, 7 Source ofNgoa ravine, 8 Ca ravine ridge – Tun ravine ridge

- Ky Son district has only one route: Cao Veu mountain

b Establishing up standard subplots

Each route is established one or two standard subplots, the method of setting

up standard subplots according to Hong Vo Van et al (2007)

The standard subplot has collected the data: coordinates, type of distribution,density, reserves, species growing together, forest cover, regeneration, and humanimpact assessment

c Data processing of fieldwork

- Map making of distribution: Regional distributions of species are plottedusing MapInfo

- Calculated density individual and density regeneration

+ The Density Calculator individual uses the formula N/ha= x 10.000(trees/ha); N is the number of trees in one hectare; n: Number of trees in a standardsubplot; So: Area of standard subplot (m2) (3-1)

+ The Density Calculator regeneration uses the formula: N/ha = (trees/ha) (3-2)Sdt: Regeneration survey area (m2); n: The number of regenerated trees wasinvestigated (HongVoVan et al., 2007)

+ Calculated Reserves

Forest reserves are calculated using the formula: M= ΣG x H x f1.3(m3); ΣG:Total basal area of a forest stand, H: Average height of the trees, f 1.3: Figure 1.3(0.483) (Hong Vo Van et al., 2007), (Forest Inventory Planning Manual, 1995)

- Handling samples and building the list of plant species growing together+ Samples collected from the field handled according to Thin Nguyen Nghia (2007).+ Identification of species names

The specimens were identified mainly based on comparative morphological

methods, mainly according to the “Flora of Vietnam” (Ho Pham Hoang, 3

volumes), Checking the species name according to “Look up the name of flora inVietnam” of Chi Vo Van (2007)

- Identify the relationship between F hodginsii and C konishii with tree layer

species commonly applied formula according to Hung Trieu Van (1994):

Number of plots with the individual appears

Total number of plots

Number of individuals of a tree species

Total number of individuals of species

Of which:

P0 is calculated according to the frequency of the survey poit

Pc is calculated according to the frequency of individualResults will be divided into 3 groups:

Group 1: Very common, including species with P0 > 30% and Pc > 7%

Group 2: Common, including species with30% ≥ P0≥ 15% and 7% ≥ Pc≥ 3%Group 3: Less common, including species with P0< 15 % and Pc< 3 %

- Method of charting vertical slice of a biome according to the method ofRichards and Davis (1934)

2.3.4 Experimental method

2.3.4.1 Research methods in morphology, tracking seasonal development

Observing, describing the morphological characteristics of parts of mature

trees that combine tracking seasonal development of F hodginsii and C konishii are

monthly for 3 consecutive years (from 2013 to 2016)

2.3.4.2 Collecting method and making microsurgical specimens

Sampling and preservation of samples in the field, microsurgery samples weremade at the Vinh University Centre for Practices and Experiments Application ofstemming techniques according to Klein R M and Klein D T (1979)

2.3.4.3 Research methodology soil

a Soil sampling method

Collecting soil samples in some regions of F hodginsii and C konishii

distributed naturally in Chau Khe commune (Con Cuong district), Nam Dungcommune (Que Phong district), Tam Hop commune (Tuong Duong district)

b Soil analyzing method

- Classifying the type of soil samples according to Chinh Tran Van (2006)

- The soil samples were analyzed at the Laboratory in the Institute of RegionalResearch and Development, Ministry of Science and Technology according to currentstandards of Vietnam (TCVN) Analytical criteria include: pHKCl, organic carbon -

OM (%), total N(%), N is easily absorbed (mg N/100g), total P (%), P is easilyabsorbed (mgP2O5/100g), total K (%), K is easily absorbed (mg K2O/100g),Absorption capacity - CEC (meq/100g), Hydrolytic acidity (meq/100g), Moisture(%), County-level components (%)

c Soil evaluating method

Soil analysis data is compared with the domestic and foreign authors' ratingscale published by Sam Do Dinh et al (2006), Siderius (1992), Anh Nguyen (2003),Dang Nguyen The et al (2007)

2.3.4.4 Propagation method

a Collecting seeds and cuttings: Selecting time to collect seeds and take cuttings

b Chemicals, breeding instruments: for sowing seeds: Viben-C, KMnO4,boiling water, sand, foam boxes, etc ; for cuttings: IBA, NAA, IAA with differentconcentrations (1,000 ppm, 1,500 ppm, and 2,000 ppm)

c Sowing seed method, cutting method: Experimental design, monitoring, andrecording data

h Processing data on cuttings: Identify indicators: survival rate, the percentage

of roots, the number of average roots, average root length of the longest one ofcuttings, the root index (Ir) for each experimental formula according to NghiaNguyen Hoang (2001)

2.3.4.5 Analysis of the essential oil

Extraction and preservation of essential oils according to the Pharmacopoeia ofVietnam (1997) Identify of the chemical composition of the oil by gaschromatography (GC) and mass spectrometry (GC/MS), confirming the constituents

were performed by comparing the spectral data obtained by MS published in theWilley/Chemstation HP library (Adams RP, 2007), conducted at the VinhUniversity Centre for Practices and Experiments.

2.3.4.6 Genetic analysis

- F hodginsii and C konishii are collected and preserved for tree bark, red and

white timber

- Sequence analysis method for nucleotide regions 18S, matK, rbcL

Analyzing genetic parameters red and white timber was conducted atDepartment of Molecular Systematics and Conservation Genetics, Institute ofEcology and Biological Resources, Science and Technology of Vietnam Includessteps: Total DNA extraction; Duplication of DNA by Thanh Nguyen Duc (2014);Data analysis (Tamura K., 2015)

Chapter 3 RESULTS AND DISCUSSION

3.1 Morphological and anatomical characteristics

Describing and adding morphological and anatomical characteristics of roots,

trunks, leaves, cones, and seeds of F hodginsii and C konishii.

3.1.1 Morphological and anatomical characteristic of F hodginsii

F hodginsii is a large evergreen trees with white and red timber, scale leaves

without petioles, leaf morphology varies There are two types of leaves: nutritionalleaves are in reverse form (4 - 5 mm) and the small reproductive leaves are scaly (2 -

3 mm) Male and female cones occur on the same plant: Male cones are egg shape orellipse, with length (4 - 5 mm), on each scales of male cones has two anthers; femalecones are spherical or near-spherical, length (1.5-1.8 cm) and width (1.5-1.8 cm), oneach of conical female has two seeds Seed length is 4- 5 mm, with wings on bothsides that are uneven The vascular bundle system of leaves, stems, and roots piledtogether in a circle, developed centrifugally

3.1.2 Morphological and anatomical characteristics of C konishii

C konishii is a large evergreen tree, white or red timber, the bark releases

white resin, shaped strip leaf, with two strands of stomata in the underside of theleaves, the leaves vary in length from 1.1 - 8 cm, width from 0.2 to 0.3 cm Male andfemale cones occur on the same plant: small male cones, oblong shapes 0.8 - 1.4 cmlong, 0.2 - 0.3 cm wide, on each male conical flakes carrying 2 anthers; female conesare conical or spherical, length (3 - 4 cm) and width (2 - 2.5 cm), on each scales offemale cone has three seeds The seeds have two wings in both sides which are quite

large, length (4-5 mm) and width (1 - 1.5 mm) The vascular bundle system of leaves,stems, and roots piled together in a circle developed centrifugally Soft tissue in thebody, thin cell wall.

3.2 Seasonal development characteristics

3.2.1 Seasonal development characteristics of F.hodginsii

In Western Nghe An BR:

F hodginsii: the cones appear in February and March, cones ripe from

December this year to early January next year, buds burst in February, March andAugust, September

C konishii: the cones appear in January and February, cones ripe in November

and December buds burst in February and March, leaves fall in October and November

According to Nguyen Duc To Luu, Philip Ian Thomas (2004) and Phan Ke Loc

et al (2007), F hodginsii cones ripe in October and November This research result

on time of ripe cones of F hodginsii in the study area shows that there is the

movement in time of ripe come from the end of December this year to early Januarynext year

3.2.2 Seasonal development characteristics of C konishii

Time of ripe cones of C konishii in the Western Nghe An BR is similar to

findings of Nguyen Duc Luu and Philip Ian Thomas (2004)

F hodginsii and C konishii are planted in forest plantation in Tay Son commune,

Ky Son district showed that: F hodginsii grows slowly, C konishii grows faster than F hodginsii In nature, when these two species reach a diameter of 0.8 m - 1.0 m or more,

in the core of the trees begins to hollow out from the root Especially C konishii, there

are many natural individuals which die without the impact of humans

3.3 Some ecological characteristics

3.3.1 Distribution characteristics of F hodginsii and C konishii

F hodginsii distributes in 20 communes belonging to 6 districts (Que Phong, Quy Chau, Tuong Duong, Ky Son, Con Cuong, and Anh Son).C konishii distributes

in 20 communes belonging to 5 districts (Que Phong, Quy Chau, Tuong Duong, Ky

Son, and Con Cuong) in Western Nghe An BR (Table 3.3 F hodginsii and C konishii have the similar distribution, appearing in the middle and high mountain

ranges divided into three main areas: the North and the Northwest; South andSouthwest and East of the biosphere reserve These conifers are distributed mostly inthe Vietnam-Laos border communes

Fokienia hodginsii usually grows near the top and top of the mountain, densely

summed, scattered or clustered around 3 - 5 individuals or mainly concentrated toform a population of 25 to 150 individuals The largest F hodginsii populations are

present at the boundary of 150 subregions with 148 subregions in Quang Phongcommune, Que Phong district (N 19025.572’ E 1040 48.522’).

C konishii usually grows from the edge of the slit to the middle of the

mountain, rarely appear at the top of the mountain, singly or in groups of about 5-7individuals or mainly concentrated to form a population from 27 to 222 individuals

C konishii population is the largest number of 222 individuals in Hach Dichcommune, Que Phong district (N 19047.012’ E 1040 51.36,8’) (Figure 3.13)

Table 3.3 Distribution of Fokienia hodginsii and Cuninghamia konishiiinWestern Nghe An Biosphere Reserve

9 Chau Hoan 1 228 228, 232 Quy Chau

16 Nam Can 1, 2 499, 500 A,C

Duong

Ban QLRPHTuongDương

Pu Mat NP

F hodginsii C.

konishii

19 Chau Khe1 787A,B,779,7

95,808

787A, B,

794, 795,

798, 813

Con Cuong

20 Luc Da1,2 796A, 805

22 Phuc Son1, 2 833, 947A - Anh Son

Notes: 1 : The new region distribution of F hodginsii

2 : The new region distribution of F hodginsii and C konishii

*: The status of forestry in the whole province according to the 2014 survey of Nghe An province (Source: North Central Forest Inventory and Planning Institute).

Figure 3.13 Distribution map of Fokienia hodginsii and Cuninghamia konishii in Western Nghe An Biosphere Reserve

3.3.2 Density, area, and reserves

3.3.2.1 The density of F hodginsii and C konishii