Rice Science, 2012, 19(1): 21−28

Copyright © 2012, China National Rice Research Institute

Published by Elsevier BV. All rights reserved

Genetic Diversity of Wild Rice Species in Yunnan Province of China

CHENG Zai-quan1, YING Fu-you1, LI Ding-qing1, YU Teng-qiong1, FU Jian1, YAN Hui-jun2,

ZHONG Qiao-fang1, ZHANG Dun-yu1, LI Wei-jiao1, HUANG Xing-qi1, 2
(1Biotechnology & Genetic Resource Research Institute, Yunnan Academy of Agricultural Sciences, Kunming 650223, China;
2
Flower Research Institute, Yunnan Academy of Agricultural Sciences, Kunming 650223, China)

Abstract: Yunnan Province of China is one of the important centers for origin and evolution of cultivated rice worldwide. Wild
rice is the ancestor of the cultivated rice. Many elite traits of wild rice have widened the genetic basis in cultivated rice.
However, many populations of wild rice species have disappeared in the past few years. Therefore, the current status of wild
rice resources should be updated and the genetic diversity of wild rice species should be examined for further germplasm
preservation and utilization. Our investigations showed that the number of natural wild rice populations declined sharply in
Yunnan Province during the past few years due to various reasons. Fortunately, one population of Oryza rufipogon, three of O.
officinalis and ten of O. granulata have been newly found in different ecological sites, which were confirmed by inter-simple
sequence repeat (ISSR) marker analysis in this study. ISSR analysis and investigation of some important traits of nutritional
values indicated that the genetic diversity of the currently existing wild rice resources in Yunnan is still rich. The
demonstration of genetic diversity of wild rice by a combined use of geographical distribution, morphological traits, nutrition
contents and ISSR markers would be helpful for the conservation and exploration of these important wild rice resources.
Key words: wild rice resource; natural distribution; genetic diversity; genetic trait

Genetic resources play an important role in
increasing yield potential in rice. As genetic diversity
among commercial cultivars decreases, it’s getting
difficult to find new genes from cultivars for futher
improvement of grain yield and quality, and sufficient
resistance to biotic or abiotic stress, which motivates
scientists to explore wild species to meet those demands
(Wang et al, 2000). Wild rice is an important gene
bank harboring lots of good traits and genes to enrich
the genetic basis of cultivars (He, 1998; Fan et al,
2000a, b; Qin et al, 2000; Zhong et al, 2000; Cai et al,
2001; Feng et al, 2002; Li et al, 2002). There are many
successful examples of dramatic increases of rice
yield with the employment of wild resources, which
contributes to the solution of the food problem for 1.3
billion people in China (Zhou and Tan, 1999). During
1926–1937, by making use of the natural hybrids of
Oryza rufipogon (wild rice) with cultivated rice, Ding
et al bred the variety Zhongshan 1 with wild rice
genes. Up to date, at least 95 varieties have been bred
using Zhongshan 1 as a parent (Li et al, 2009).
Another example is that YUAN Longping and his
colleagues led national effort to develop hybrid rice
with male sterile genes from O. rufipogon (Pang,
1998). The hybrid rice has tremendousely increased
grain yield of rice and made great contribution to
world food production. The gene WBB1 responsible
Received: 29 November 2010; Accepted: 23 February 2011
Corresponding author: HUANG Xing-qi (xingqih@public.km.yn.cn)
for resistance to rice bacterial blight in the entire life
of rice was derived from O. rufipogon as well (Zhang,
2002). Many other reports have demonstrated various
good traits derived from wild resources to improve
rice cultivars (Pang, 1998; Lu et al, 2002; Yang,
2003).
Wild rice distributes in all continents except Europe,
North America and Antarctic areas in the world,
mostly in tropical and sub-tropical areas of Asian.
China, as one of the important origin countries of wild
rice, has three major wild rice species: O. rufipogon
Griff. (A′A′, 2n = 24), O. granulate Baill (GG, 2n = 24)
and O. Officinalis Wall (CC, 2n = 24) (Zhang, 2002).
Yunnan Province is located in the southwest China at
a latitude from 21°09′32′′ to 29°15′04′′ N and a
longitude from 97°31′39′′ to 106°11′47′′ E, with
altitude varying from 76 m above sea level to 6740 m.
The mountains and rivers run from north to south
across the province, which forms eight climate zones
with unique topography and biological resources.
Yunnan Province has been considered as one of the
precious green treasures on the earth, known as the
‘plant and wildlife kingdom’, with great biodiversity
(Fan et al, 2000b). Many wild rice populations have
been found in the past few decades since the first O.
officinalis was found in Jinghong of Yunnan in 1936
by WANG Qiyuan (Cheng et al, 2004). According to
some previous reports, wild rice populations were
identified in pools, river sides, bamboo fields, banana
22
tree fields, rubber tree fields, shrub fields and forest
fields from the altitude of 400 m to 1000 m in the
southern five prefectures of Yunnan. The number of
wild rice species and ecological types (populations) in
Yunnan Province were the highest among all related
provinces of China (Gao et al, 2000). Each wild rice
species has many subtypes or ecological populations
in Yunnan Province, which can be used for breeding
rice cultivars with high yield and multiple resistances.
Many scientists reach an agreement that Yunnan wild
rice is one of the most important parts of wild rice
resources in China because all the three wild species
in China have been found in Yunnan, and each species
has a lot of ecotypes. Due to the increasing interference of
human being activities, the proper ecological
environments for many plants including wild rice have
been changed or damaged seriously. Many recent
reports have shown that wild rice resources were
consistently reduced and some natural populations even
disappeared in Yunnan since the 1990s. Therefore,
wild rice resources need to be protected and conserved
urgently (Fan et al, 2000b; Gao et al, 2000). However,
several issues have occurred to the rice community in
Yunna Province, such as the possibility to find new
wild rice populations, recent changes of wild rice
resources, important traits in the wild rice species for
cultivar improvement, the genetic diversity in the wild
rice resources nowadays, and molecular analysis and
comparison of morphological traits and some important
nutritional contents of seeds. In order to answer these
questions, we carried out the following studies in the
past few years for the genetic diversity of wild rice
resources in situ in Yunnan Province of China.
MATERIALS AND METHODS
Field investigation
Comprehensive investigation of wild rice resources in
Yunnan
From 2001 to 2006, investigation was conducted in
Yunnan to find new populations of wild rice resources
and to confirm if the wild rice plants in those
populations identified previously still exist naturally.
Some plants of the presently existing wild rice
populations were grown in a relocation base. The
conditions for growing the wild rice plants were
similar to cultivated rice. Rice cultivars as control
were also planted about 3 km far away from the wild
rice relocation base. Important characteristics of some
wild rice plants of nine ecotypes were investigated
during the whole growth period for three years.
Rice Science, Vol. 19, No. 1, 2012
Morphological investigation of wild rice in Yunnan
Important traits of 10 plants sampled from each
wild rice ecotype were scored and recorded in the
relocation base and in the original populations. The
relocation bases of all the three wild rice species have
similar climate and environmental conditions.
Evaluation of agronomic traits
Resistance to diseases and abiotic stress in wild rice
of Yunnan
The methods reported by Feng et al (2002) and
Yoshimura et al (1998) were used to evaluate the
resistance to blast and bacterial blight, respectively.
The methods reported by Fan et al (2000a, b) were
adapted to evaluate the resistances to abiotic stress of
drought and chilling.
Protein and amylose contents in husked wild rice
seeds
Eight ecotypes of wild rice species (Table 1) and six
cultivated rice (O. sativa) (Table 2) were analyzed.
Seeds of wild rice were collected from their original
populations or the wild rice protection populations in
Jinghong. The cultivated rice varieties Yunhui 290 and
Dianchao 9 belong to indica rice, Hexi 35 and Yuyou
1 belong to japonica rice, Xiangyaxiannuo belongs to
indica glutinous rice and X-Jingdaonuo belongs to
japonica glutinous rice. Seeds of each cultivated rice
were collected from the similar regions of wild rice
and were husked. Total protein content in the husked
grains was measured by the method of Zhen et al
(1997). Amylose content was determined according to
the methods of Cheng et al (2004) and Zhang et al
(1996). Each assay was repeated three times and data
were presented as the mean ± SD.
Inter-simple sequence repeat (ISSR) analysis
Plant materials
Five sub-populations of Yuanjiang O. rufipogon and
four sub-populations or types of Jinghong from Yunnan;
four of O. officinalis (two populations from Jinghong,
wide leaf type and normal type, one from Mengding,
and one from Gengma); 16 of O. granulata from different
ecological regions were used for ISSR analysis. Some
of them were newly found populations.
Total genomic DNA extraction
SDS method was used to extract total genomic
DNA of rice. The procedure of genomic DNA extraction
of O. granulata was modified slightly from the SDS
method (Sun et al, 1997).
ISSR analysis
A total of 22 ISSR primers were ordered from the
Shanghai Sangon Company (China). After screening,
CHENG Zai-quan, et al. Genetic Diversity of Wild Rice Species in Yunnan Province of China 23

Table 1. Summary of important characteristics of representative populations of three wild rice species in Yunnan, China.
Species Type Population Important characteristics
O. rufipogon Erect type Banna Strong and board-spectrum resistance to blast, thick and erect leaf blade, relatively erect
stem, high-tilling, big panicle, high seed-setting rate, good grain quality, big anther,
strong growth ability
Red-awn type Banna Moderate resistance to blast, strong resistance to bacterial blight, thick and erect leaf
blade, extremely high-tilling, big panicle, high seed-setting rate, good grain quality, big
anther, strong growth ability
Yuanjiang Moderate resistance to blast, strong resistance to bacterial blight, thick and erect leaf
blade, extremely high-tilling, extreme tolerance to chilling, drought tolerance, good
grain quality, big anther, strong growth ability
O. officinalis Gengma Moderate resistance to blast, resistance to bacterial blight, resistance to stemborer and
planthopper, exellent grain quality, big panicle, thick rhizome, strong growth ability,
outward stigma, extreme tolerance to chilling
Mengding Resistance to blast, bacterial blight, stemborer and planthopper, high seed setting rate, good
grain quality, big panicle, thick rhizome, strong growth ability, extreme tolerance to
chilling
Jinghong Similar to O. officinalis from Gengma, except for extremely broad leaf blade and chilling
tolerance
O. granulata Banna Extreme resistance (immunity) to bacterial blight, big and outward stigma, thich
rhizome, drought tolerance, reproductive growth period is extremely long (nor light
sensitiveness), wide adaptability
Mengding Similar to O. granulata from Banna with good grain quality
The underlined are the traits investigated newly. The traits in bold are the most valuable for improvement of rice cultivars.

Table 2. Seeds of different Yunnan wild rice species or types. Table 3. ISSR primers used in analysis of genetic diversity.

Species Type Collected site Collected date Primer Primer sequence

ISSR1 BDBCACACACACACA
O. rufipogon Yuanye Yuanjiang Nov. 2001 ISSR2 VHVGTGTGTGTGTGTGT
Yuanye Jinghong Nov. 2001 ISSR3 DBDGAGAGAGAGAGAGA
Jingye Jinghong Nov. 2001 ISSR4 GCACACACAC
O. officinalis Yaoye I Jinghong Nov. 2001 ISSR5 CTCTCTCTCTCTCTCTRG
Yaoye III Kunming Nov. 2001 ISSR6 CTCTCTCTCTCTCTCTRC
Yaoye I Jinghong Nov. 2002 ISSR7 CCCGTGTGTGTGTGT
Yaoye II Zhedian Nov. 2001 ISSR8 GSGGTGTGTGTGTGT
Mengding ISSR9 CSCGAGAGAGAGAGA
Yaoye IIIA Gengma Nov. 2001 ISSR10 GCWGAGAGAGAGAGAG
O. granulata Youye Jinghong Nov. 2001 ISSR11 CCAGTGGTGGTGGTG
Youye Mengding Nov. 2001 ISSR12 GCGACACACACACACA
O. sativa Yunhui 290 Jinghong Sept. 2001
Diancha 9 Jinghong Sept. 2001
Yuyou 1 Yuxi Sept. 2001
Molecular analysis of genetic diversity of wild rice
Hexi 35 Yuxi Sept. 2001
Xiangyaxiannuo Jinghong Sept. 2001 resources in Yunnan
X-Jingdaonuo Yuxi Sept. 2001
The materials were the representatives of different
populations in Yunnan. The molecular analysis method
12 primers (Table 3) were selected for genetic was similar to the above description.
diversity analysis to all samples of the three wild rice
species (Chen et al, 2002). RESULTS
PCR conditions were described according to the
method of Wu et al (2004). Data analysis including New populations of wild rice identified in Yunnan
percentage of polymorphic bands and Nei’s gene
Our comprehensive investigation identified several
diversity was conducted using POPGENE (version
new wild rice populations. One population of O.
1.32) software. Jaccard similarity coefficients were
rufipogon was found in Jinghong, belonging to
calculated using NTSYS-pc (version 2.0) (Numerical
Taxonomy System, Rohlf, 1993). Genetic relationship Jinghong type of O. rufipogon. The plants in the new
among samples was analyzed using the unweighted population were different from those in other
pair group method (UPGMA) (Sneath and Sokal, 1973). ecological populations or types of O. rufipogon with
24 Rice Science, Vol. 19, No. 1, 2012

resistance to insects or diseases. The main differences
were shown in wider and longer leaves, bigger
panicles and higher seed-setting rate, and the blast
resistance was stronger in Jinhong type than in other
types of O. rufipogon. Two new populations of O.
officinalis were found at Genma and Menghai,
respectively. They were the only naturally existing O.
officinalis popuplations in Yunnan. Ten new
populations of O. granulata were found in some
banana fields, riversides and forest fields in Jinghong
and Simao of Yunnan.
All the newly found populations of wild rice were
ecological populations differred from the other
previously found populations. The ISSR analysis showed
some different traits and genetic backgrounds of these
newly found populations (Fig. 1).
Comparison of the population number of wild rice
between new and existent populations
O. rufipogon
Our investigation showed that 23 previously found
populations of O. rufipogon had no wild rice plants in
Yunnan at present. This fact indicates that in the past
20 years, wild rice in situ became extinct at a
surprisingly high speed that almost one population (in
one ecological site) of O. rufipogon disappeared each
year in Yunnan. One population of O. rufipogon
recently found at Mandiu of Jinghong has some wild
rice plants, and another population of O. rufipogon
previously found at Yuanjiang also has some wild rice
plants, but they differentiated into five sub-populations.
Actually, it was found that there were many plants in
Yuanjiang O. rufipogon population, although the plant
number was once reduced in the past 20 years. The
plant number of O. rufipogon in Yuanjiang was only
15−20 in 2003, but gradually increased to 380–400 in
2006 after establishment of protection base around the
Yuanjiang population.
O. officinalis
O. officinalis distributed over five counties of three
prefectures in Yunnan. The total population number of
the identified O. officinalis was 14 (Cheng et al, 2004).
Our investigation showed that 12 of the 14
populations had no plants at their original sites now. It
is surprising that in the past 20 years, almost one
natural population of O. officinalis disappeared in
every two years in Yunnan. Fortunately, we found two
populations of O. officinalis that were never recorded
before, one in Gengma and the other in Menghai. The
plant number of O. officinalis in Gengma was only
5–6 in 2002. After construction of protection base at
that site, the plant number recovered gradually, and
reached 40–50 in 2006.
O. granulata
O. granulata distributed widely in Yunnan. So far,

Fig. 1. Genetic backgrounds revealed by ISSR-marker based analysis of different populations of three wild rice species in Yunnan, China.
A, O. rufipogon. 1, Dongxiang from Jiangxi Province; 2, Mandiu, Jinghong (1); 3, Mandiu, Jinghong (2); 4, Purple-stem of Jinghong; 5,
Green-stem of Jinghong; 6, Yuanjiang (1); 7, Yuanjiang (2); 8, Yuanjiang (3); 9, Yuanjiang (4); 10, Yuanjiang (5); 11, O. longistaminata; 12, Yuyou 1;
13, Hexi 35; 14, NC; 15, Dianlong 201; M, M2000 marker. B, O. officinalis. 1, Jinghong 1; 2, Jinghong 2; 3, Menghai; 4, Mengding; 5, Wide leaf
type. C, O. granulata. 1, Jinghongxi; 2, Zhengkang; 3, Mengbeixin; 4, Simao; 5, Erhe; 6, Chishuiguan; 7, Mengkuang; M, M2000 marker. The
underlined are newly found populations of wild rice in Yunnan Province, and others are wild rice reported before and the cultivated rice varieties. The
numbers in the parenthesis present the subpopulation of sample plants from different wild rice growing fields in the same original site.
CHENG Zai-quan, et al. Genetic Diversity of Wild Rice Species in Yunnan Province of China
some O. granulata populations have quite many plants
scattering naturally in large areas up to several hectares.
This was very special for O. granulata in comparison
to small population size naturally for O. rufipogon and
O. officinalis. Data of investigation since 1950s
indicated that O. granulata including 122 populations
distributed in 20 counties of 7 prefectures in Yunnan.
Ten populations of O. granulata have been recently
found from year 2001 to 2006. Our investigation
concluded that 85 of the previously found populations
of O. granulata have no plants at their original sites
presently. In other words, O. granulata permanently
disappeared at a speed of 3–4 populations each year in
the past two or three decades in Yunnan. At present,
only 37 populations of O. granulata have plants at
their original sites, mainly at Simao and Lincang
prefectures in Yunnan (Fig. 2).
Wild rice resources need urgent protection in Yunnan
Our investigations suggested that the extinction of
wild rice resources in Yunnan urgently needs public
attention. Take O. granulata as an example, in the past
three decades, the extinction speed of O. granulata
populations have kept increasing. Consequently, 85 of
the 122 previously identified populations have disappeared.
Among the 37 existing populations of O. granulata at
their original sites currently, 28 (76%) are approaching
to the edge of extinction. The environmental conditions
are becoming more unfavourable for the survival of
these 28 populations of O. ganulata. Plant number in
each of these 28 populations is gradually reducing.
Unless some effective ways are taken to protect these
populations, O. granulata would disappear sooner or
later in Yunnan. Only 9 populations of O. granulata
are relatively safe at present, because their original
sites are in steep mountains that are free from the
interference of human beings and animals.
In general, the populations and the plant number in
each population of Yunnan wild rice have decreased
sharply in the past 20 years, especially for O. rufipogon
Fig. 2. Comparison of wild rice populations at different periods.
25
and O. officinalis. It is surprising that the extinction
rate of wild rice in Yunnan is extraordinarily high.
Genetic diversity of wild rice resources assessed by
phenotypic traits in Yunnan
Prior to this study, no systematic investigation was
conducted to phenotypically characterize the wild rice
species, especially for growth-developmental characteristics
during the whole life period. The important or special
traits of the three wild rice species are summarized in
Table 1. It was found that each wild rice species had
unique or important traits that were not observed in
other species and cultivars. For example, O. rufipogon
from Yuanjiang displayed extremely strong tolerance
to chilling. The plants of O. officinalis from Gengma
had the highest seed setting rate (82.0% ± 1.2%)
among all the wild rice species. O. ganulata had
extremely strong tolerance to drought with a long
reproduction period from April to November,
indicating it is photo-period insensitive.
Moreover, all the three wild rice species shared
some characteristics with cultivated rice. For example,
all the three wild species had tolerance to bacterial
blight and long outward stigma. Most of the plants of
different wild rice species maintained their phenotypic
traits in the relocation base after being transplanted
from their own original populations. This observation
suggests that most of the traits are genetically
controlled. However, some traits such as plant height
changed. For example, the plant height of O.
officinalis plants under shrub reached 4 m in Gengma,
but only 1.5–2.0 m at the relocation base. The plant
height of an O. granulata population reached 80–90
cm in Dongwo, but only 60 cm at the relocation base.
The average protein contents in the husked seeds of
O. rufipogon, O. granulata and O. officinalis were
14.47% ± 0.64%, 16.28% ± 2.05% and 15.30% ± 0.57%,
respectively (Fig. 3). O. officinalis from Gengma had
the highest protein content of 19.3%. The seed protein
content was significantly higher in wild rice species
than in O. sativa, suggesting greater nutritional value
of wild rice.
Amylose contents ranged from 8.8% to 13.0%
among the samples of Yunnan wild rice, with an
average of 12.0% ± 0.38%, 9.70% ± 1.40%, and
11.28% ± 0.43%, respectively for O. rufipogon, O.
officinalis and O. granulata (Fig. 4). All the three wild
rice species had significantly lower amylose content
than O. sativa. Generally speaking, rice with low
amylose content is favored for steamly cooked rice
because of softness and taste feeling. This observation
indicates that wild rice may have genes controlling
lower amylose content, which would be useful to
26 Rice Science, Vol. 19, No. 1, 2012

populations of O. rufipogon conserved in the relocation

base. The five populations of O. rufipogon were
highly diverse, expressed by Nei’s value of 0.26
among different populations and 0.13 among the
Protein content (%)

plants inside the populations (Table 4). The ISSR

markers generated 80 polymorphic bands with a
percentage of polymorphic bands of 64.52%. The genetic
distance (I) was 0.37 among different populations and
0.19 among the plants inside the populations. The
genetic distance of O. rufipogon was high between
Yuanjiang and Jinghong type. The differences of these
Fig. 3. Average protein content in the husked seeds of three wild types of O. rufipogon were similar in the morphological
rice species and O. sativa. and molecular analysis.
AA, O. sativa; A′A′, O. rufipogon; CC, O. officinalis; GG, O.
granulate. Genetic diversity of O. officinalis in Yunnan
The genetic variation among different populations was
20
higher than the variation among different plants inside
16
15.96±1.16
a population of O. officinalis. The genetic variation
coefficient (h = 0.22) among populations of O.
Amylose content (%)
Percentage (%)

12.0±0.38
12
9.70±1.4
11.28±0.43
officinalis was lower than that of O. rufipogon.

8 Genetic diversity of O. granulata in Yunnan

3.88±1.03
4
0
AA(g) AA
Fig. 4. Average amylose content in the husked seeds of three wild
rice species and O. sativa.
AA, O. sativa; A′A′, O. rufipogon; CC, O. officinalis; GG, O.
granulate. AA(g) means glutinous rice of O. sativa.
improve rice quality.
In general, most of the wild rice species found in
Yunnan have many special and important traits, such
as strong resistance and good quality which are very
useful for rice cultivar improvement.
Genetic diversity of wild rice resources assessed by
ISSR analysis in Yunnan
Genetic diversity of O. rufipogon in Yunnan
A total of 12 ISSR primers were identified
polymorphic among the sampled entries of O.
rufipogon (Table 3). There were two populations of O.
rufipogon naturally existing in Yunnan, and three
For 16 populations, 135 bands were resulted from
the PCR with 12 ISSR primers, with an average of
11.2 bands per primer. Among 135 bands, 93 were
A'A' CC GG polymorphic with the percentage of polymorphic bands
of 68.8%. The lengths of amplified bands ranged from
100 to 2 000 bp. In addition, the observed average
number of alleles (Na) was 1.81, the mean Nei’s gene
diversity (h) was 0.33 and the mean Shannon
information index (I) was 0.43.
Within the population Banna, 105 bands were
resulted from the PCR with 10 ISSR primers (ISSR1–
ISSR10). However, only 27 bands were found to be
polymorphic with the percentage of polymorphic bands
of 25.7%. Similarly, in the population Mending, 78
bands were produced after PCR with 8 ISSR primers
(ISSR1, ISSR2, ISSR4, ISSR5, ISSR6, ISSR9, ISSR10,
ISSR12). Among 78 bands, only 17 were polymorphic,
showing the percentage of polymorphic bands of 21.8%.
A cluster analysis was used to generate two
dendrograms based on the Jaccard coefficients of
ISSR markers among 14 populations of O. granutula

Table 4. Genetic diversity of two wild rice species revealed by the ISSR marker analysis.
No. of ISSR No. of ISSR
Species Range PPB of ISSR Na Ne h I
bands polymorphic bands
O. rufipogon Among 5 populations 124 80 64.52 1.66 1.45 0.26 0.37
Within population plants 75 25 33.33 1.34 1.23 0.13 0.19
O. granulata Among 16 populations 135 93 68.84 1.81 1.52 0.33 0.43
Within Population Banna 105 27 25.71 1.25 1.15 0.09 0.14
Within Population Mending 78 17 21.79 1.22 1.11 0.07 0.10
PPB, Percentage of polymorphic bands; Na, Observed number of alleles; Ne, Effective number of alleles; h, Nei’s gene diversity; I, Shannon
information index.
CHENG Zai-quan, et al. Genetic Diversity of Wild Rice Species in Yunnan Province of China 27

(Fig. 5), and a cluster analysis of ISSR markers within Ind1

Ind2
population Banna (Fig. 6). Ind3
Ind4
The genetic diversity was high among 16 Ind5
Ind17
populations, but low within any population of O. Ind18
Ind19
granulata in Yunnan. The population number was Ind20
Ind7
larger in O. granulata than in O. rufipogon. The Ind6
genetic diversity (h) and distance (I) of O. granulata Ind8
Ind9
populations were higher than those of O. rufipogon Ind10
Ind16
populations. However, the genetic diversity (h) and Ind21
Ind22
distance (I) within an O. granulata population were Ind23
Ind24
lower than those within an O. rufipogon population. In Ind11
Ind12
general, O. granulata was genetically more diversified Ind13
Ind14
than either O. rufipogon or O. officinalis. Ind15

0.85 0.90 0.95 1.00

DISCUSSION Coefficient (Jaccard)
Fig. 5. Dendrogram of 14 populations of O. granutula based on the
Currently, wild rice O. rufipogon only naturally results of ISSR marker analysis by UPGMA method.
exists in Yuanjiang and Jinghong in Yunnan province
and most of the plants within a population are
genetically diversified. Fortunately, we had collected
some plants from three populations that have naturally
disappeared and preserved them in the relocation base.
Large genetic distance has been found among five
populations of O. rufipogon. It is surprising to discover a
very large genetic distance between Yuanjiang and
Jinghong types of O. rufipogon, but these two types
have a close genetic relationship with Dongxiang type
of O. rufipogon from Jiangxi Province, China. In
addition, there is a rich genetic variation among
different plants in the currently existing populations ex
situ. It is a pity that some populations of O. rufipogon
in Yunnan have disappeared and the genetic diversity in Fig. 6. Dendrogram of population Banna of O. granutula based on
ISSR marker analysis by using UPGMA method.
those populations have lost forever. Fortunately, the
genetic diversity among the currently existing O.
rufipogon is still very high among the five populations Luckily, there is a certain degree of genetic diversity
and the plants of each population. This conclusion among different populations and different plants inside
receives a support from the substantial variation for each ex situ population of O. officinalis preserved and
morphological characteristics and growth-development protected in the relocation base and in situ population
traits among the currently existing populations of O. in original sites (Cheng et al, 2004).
rufipogon in Yunnan (unpblished data). Even though the number of natural populations for
It is noticed that the genetic diversity of O. O. granulata resource have seriously decreased in the
officinalis has dramatically decreased because many past (Qian et al, 2001), there are 37 naturally ecological
naturally ecological populations have disappeared populations at present in Yunnan. In addition, there are
during last two or three decades (Chen et al, 2002). other three ex situ populations collected and preserved
There are two naturally ecological populations of O. in the relocation base. O. granulata has more
officinalis, and other two ecological populations populations than either O. rufipogon or O. officinalis.
collected and preserved ex situ in the relocation base. The results from molecular marker analysis showed
Great amount of variation in plant height, leaf size, that O. granulata had the most genetic diversity
stem size and seed-setting rate have been found among the three wild rice species in Yunnan.
among the four populations. Results from molecular
marker analysis of ISSR showed a great amount of ACKNOWLEDGEMENTS
genetic diversity. Relatively speaking, the genetic
diversity in O. officinalis is lower than in O. rufipogon. This work was supported by the Yunnan Natural
28
Science Foundation Project (Grant No. 2008C004);
the Natural Science Foundation of China (Grant No.
31160067), and Special Fund for Agro-scientific
Research in the Public Interest (Grant No. 201003021).
We also thank Dr. YAN Wengui from Rice Center of
United States Department of Agriculture (USDA) for
his proofreading of this paper.
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