Journal of Genetics and Genomics (Formerly Acta Genetica Sinica) September 2007, 34(9): 851-860

851

Research Article

Genetic Diversity of Maize (Zea mays L.) Landraces from Southwest China Based on SSR Data
Qilun Yao 1, 2, Kecheng Yang 1, , Guangtang Pan 1, Tingzhao Rong 1
1. Maize Research Institute, Sichuan Agricultural University, Ya 625014, China; an 2. Department of Life Science, Yangtze Normal College, Fuling 408003, China

Abstract: Genetic diversity of 54 maize landraces from southwest China was tested using bulk DNA samples and 42 microsatellite (SSR) loci distributed on 10 chromosomes of maize. A total of 256 alleles were detected among the landraces. At each locus, the number of alleles varied from 2 to 9, with an average of 6.1. On the basis of the genetic similarity coefficients, clustering analysis separated the landraces into four groups. The landraces collected from the same region were mostly grouped together. To reveal the genetic structure and genetic diversity within landraces, 165 individuals from 11 landraces were analyzed. Individual DNA samples proved to be superior to bulk DNA samples in identifying genetic diversity of landraces. A total of 330 alleles were detected in the 11 landraces. According to the results of the individual DNA sampling analysis, estimates of the mean number of alleles A, the effective allelic number Ae, the observed heterozygosity Ho, and expected heterozygosity He were 7.86, 3.90, 0.69, and 0.37, respectively. An obvious genetic deviation from Hardy-Weinberg expectation was observed both among and within landraces and a considerable genetic variation was revealed within rather than among landraces. In addition, genetic diversity of landraces was greater in Sichuan than in the other three regions. It can be concluded that maize landraces in southwest China were initially introduced to Sichuan and from there to adjacent areas. Keywords: genetic diversity; maize landraces; SSR

Modern farming techniques have greatly increased production levels, but they have also led to greater genetic uniformity. As farmers switched to improved crop varieties on a large scale, they bandoned cultivation of landraces, leading to disapperance of landraces for the fact of not being well preserved [1]. Genetic diversity is the basic portion of biological diversity and is the base of species diversity. Genetic diversity of maize (Zea mays L.) plays a key role for future breeding progress. For several decades, maize breeders have focused on short-term breeding.

This has resulted in the development of a narrow genetic base for commercial maize hybrids. Most hybrids grown in the U.S. are derived from 68 inbred lines [2]. In China, the parenthood of 91.6% hybrids consists of about 20 elite inbred lines [3]. Although genetic gains for maize seem not to have diminished, continuous selection may have narrowed the genetic variation in hybrid germplasm. With such a restrictive base, maize may not contain all the desirable and favorable alleles to maintain selection progress.

Received: 20061122; Accepted: 20061228 This work was supported by the National High Technology Research and Development Program of China (No. 2004BA525B04), Changjiang Scholars and Innovative Research Team in University (No. IRTO453). Corresponding author. E-mail: yangkc2006@yahoo.com.cn www.jgenetgenomics.org

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More than 15,000 accessions of maize have been collected in China, and about 90% are landraces. As Chinese maize breeders use less than 3% of the total landrace germplasm available, the investigation and utilization on the landrace germplasm is limited [4]. In southwestern China, maize landraces are distributed intensively in Sichuan, Chongqing, Guizhou, and Yunnan Provinces, where 3,340 landraces can be accessed to the National Germplasm Bank [5]. According to a field survey from 2003 to 2005, farmer-saved maize landraces are still grown on mountain areas, mainly due to local adaptation of landraces and the economic factors that limit the adoption of commercial hybrids (unpublished data). These original landraces represent a vast array of germplasm, which can be used in maize breeding for improving disease and pest resistance, nutritional quality, and other traits of interest. The development of molecular markers provides the repertoire of assessing genetic diversity at the DNA level in plant species [6]. In particular, SSR makers are potential for large-scale DNA fingerprinting of maize genotypes due to the high level of polymorphism detected [7], automated analysis systems [8] and high accuracy and repeatability [9]. However, genetic diversity within and among maize landraces using DNA markers is not well studied. The objectives of this study were 1) to investigate genetic diversity of maize landraces in southwestern China; 2) to compare the results of genetic diversity from analyses of bulk DNA samples and individual DNA samples; and 3) to reveal the genetic structure of landraces.

individual plants to analyze genetic diversity of landraces. On the basis of their geographic distributions (Fig. 1), 11 landraces, each of which contained 15 individual plants, were used to study genetic variability within a landrace.

Fig. 1 Geographical distribution of the 11 landraces sampled from 4 regions in southwestern China

1.2

SSR analysis

Genomic DNA was isolated from the third fresh leaf following CTAB procedure described by Scott [10] with minor modifications. PCR amplification was performed in a PTC-220 thermal cycler (Surplus Lab Inc., Michigan, USA) programmed at 35 cycles of 1 min at 95, 2 min at 55, and 2 min at 72, followed by a 10 min extension at 72. The PCR amplification products were separated on a 6% (w/v) denatured polyacrylamide gel and visualized by silver staining. 1.3 SSRs data scoring and analysis

1
1.1

Materials and Methods

Plant materials

Fifty-four maize landraces from Sichuan, Chongqing, Guizhou, and Yunnan Provinces were used in this study (Table 1). They were planted in Plant Garden in Maize Research Institute of Sichuan Agricultural University at Ya of Sichuan Province, China. an Genomic DNA was isolated from a bulk sample of 15

The SSR bands were scored as present (1) or absent (0), each of which was treated as an independent character. Genetic diversity analysis was conducted
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Table 1
No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54

The sources of 54 maize landraces in southwest China

Landrace DP-01 DP-02 DP-03 DP-04 DP-05 DP-07 DP-08 DP-09 DP-67 DP-68 DP-11 DP-12 DP-13 DP-14 DP-15 DP-17 DP-18 DP-19 DP-20 DP-21 DP-23 DP-24 DP-25 DP-26 DP-27 DP-28 DP-29 DP-31 DP-32 DP-35 DP-39 DP-36 DP-38 DP-37 DP-40 DP-42 DP-44 DP-45 DP-48 DP-50 DP-52 DP-53 DP-54 DP-55 DP-56 DP-57 DP-58 DP-59 DP-62 DP-64 DP-60 DP-61 DP-65 DP-66 Origin (County, Province) Wengan, Guizhou Renhuai, Guizhou Bijie, Guizhou Shiqian, Guizhou Panxian, Guizhou Ziyun, Guizhou Wangmo, Guizhou Hanyuan, Sichuan Hanyuan, Sichuan Hanyuan, Sichuan Xiaojin, Sichuan Wenchuan, Sichuan Danba, Sichuan Leibo, Sichuan Leimaping, Sichuan Linshui, Sichuan Dazhu, Sichuan Nanchong, Sichuan Yilong, Sichuan Gangyuan, Sichuan Junlian, Sichuan Junlian, Sichuan Junlian, Sichuan Junlian, Sichuan Pingshan, Sichuan Pingshan, Sichuan Pingshan, Sichuan Xuyong, Sichuan Qingchen, Sichuan Neijiang, Sichuan Neijiang, Sichuan Pengxian, Sichuan Ganzi, Sichuan Jiangbei, Chongqing Fengjie, Chongqing Wushan, Chongqing Fengdu, Chongqing Fengdu, Chongqing Fengdu, Chongqing Fengdu, Chongqing Fengdu, Chongqing Shizhu, Chongqing Shizhu, Chongqing Shizhu, Chongqing Xiushan, Chongqing Xiushan, Chongqing Wanzhou, Chongqing Jiangchuan, Yunnan Jiangchuan, Yunnan Jiangchuan, Yunnan Chengjiang, Yunnan Hongta, Yunnan Wuding, Yunnan Wuding, Yunnan Germplasm characteristics description Intermediate maturing, yellow flint grain type Late maturing, white flint grain type with long ear Intermediate maturing, white flint grain type Intermediate maturing, white flint grain type with big kernel Intermediate maturing, yellow semi-dent grain type Early maturing, white dent grain type with short stalk Late maturing, yellow semi-dent grain type with big ear Intermediate maturing, white dent grain type with long ear Intermediate maturing, yellow semi- dent grain type Late maturing, yellow flint grain type with long stalk Early maturing, yellow flint grain type with short stalk Intermediate maturing, white flint grain type with big ear Early maturing, yellow semi-dent grain type with short stalk Intermediate maturing, white dent grain type with short stalk Intermediate maturing, yellow flint grain type with long stalk Intermediate maturing, yellow semi-dent grain type Intermediate maturing, white flint grain type with big ear Intermediate maturing, yellow flint grain type with long ear Intermediate maturing, yellow flint grain type with big ear Early maturing, white semi-dent grain type with big kernel Intermediate maturing, white flint grain type Intermediate maturing, red flint grain type Late maturing, blue dent grain type with big kernel Intermediate maturing, white semi-dent grain type Early maturing, white flint grain type with short stalk Intermediate maturing, yellow flint grain type with big kernel Late maturing, white flint grain type with long stalk Intermediate maturing, white flint grain type with long stalk Intermediate maturing, yellow semi-dent grain type Intermediate maturing, yellow semi-dent grain type Late maturing, yellow semi-dent grain type with big ear Intermediate maturing, yellow flint grain type Early maturing, white flint grain type with big kernel Intermediate maturing, yellow flint grain type Intermediate maturing, purple flint grain type with big kernel Late maturing, yellow flint grain type with big ear Intermediate maturing, yellow semi-dent grain type Late maturing, white semi-dent grain type with big ear Intermediate maturing, white semi-dent grain type with big ear Late maturing, yellow semi-dent grain type with long ear Intermediate maturing, white flint grain type with short stalk Late maturing, white flint grain type with big ear Late maturing, blue semi-dent grain type with big kernel Intermediate maturing, yellow dent grain type Late maturing, yellow dent grain type with long ear Late maturing, white waxy with long stalk Late maturing, white waxy with long stalk and big kernel Late maturing, white semi-dent grain type with big ear Late maturing, yellow semi-dent grain type with long ear Late maturing, white waxy with long stalk and small kernel Intermediate maturing, white dent grain type with long stalk Late maturing, yellow dent grain type with long ear Late maturing, yellow dent grain type with big kernel Late maturing, purple dent grain type with big kernel

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on the basis of the scores. The statistical methods and formulae used are described below: 1) The index of genetic similarity (GS): GS = 2Nij/(Ni + Nj), where Nij is the number of SSR alleles common to landraces i and j, while Ni and Nj are the total numbers of SSR alleles observed for landraces i and j, respectively. The dendrogram were constructed by the unweighted pair-group method with arithmetic mean (UPGMA) clustering using the computer software NTSYS-pc version 2.10. 2) The mean number of alleles A: A=

Fis are 0. Fst, ranging from 0 to 1, is an estimate of gene differentiation between landraces, which represents genetic variation among landraces [11, 13]. Fst is 0 if there was no genetic variation among landraces. 7) Genetic distances D between each pair of landraces were estimated by the modified Rogers distance as follows: D =
X Y

1 n

i =1 j =1

1 X Y ( p ij p ij ) 2 , 2

where pij and qij are the frequencies of ith allele at jth allele in landraces X and Y, respectively. All the parameters were computed using the POPGENE software version 1.2 (Department of Renewable Resources, University of Alberta, Edmonton, Canada).

i =1

Ai / n ,

where Ai is the number of alleles at ith allele. 3) The effective allelic number
Ae =

Ae:

A
i =1

ei

/n=

(1 / q
i =1 j =1

2 j)/

n , where Aei is the

2
2.1

Results
Genetic diversity among 54 maize landraces for bulk DNA samples

effective allelic number at ith allele, and qj the frequency of the jth allele. 4) Ho is the observed heterozygosity:
Ho =

H
i =1

oi

/n=

(1
i =1 j =1

mi

2 q ij ) /

n , where Hoi rep-

ressents the observed heterozygosity of the ith allele, and qij is the frequency of the jth homozygous allele at ith allele. 5) The expected heterozygosity, also known as index of gene diversity
[11]

: He =

H
i =1

/n=

q
(1
i =1 j =1

mi

Using 42 SSR loci distributed on 10 chromosomes of maize, a total of 256 alleles with 96.8% polymorphism were detected in the bulked DNA samples from 54 landraces. For each SSR locus, the number of alleles ranged from 2 to 9 corresponding to an average of 6.1 per locus, revealing a high level of genetic diversity of maize landraces in southwest China. The dendrogram based on SSR markers distinguished 54 landraces, which were distinctly clustered into four groups (Fig. 2). The biggest group (Group) consisted of 24, 10, and 6 landraces from Sichuan, Chongqing, and Guizhou, respectively. Groupincluded 8 landraces, of which 7 originated from Yunnan. Five landraces from the 4 regions were clustered into Group . In addition, landrace DP-05 was less related to other landraces genetically, and formed Group . Comparison of the landraces within the same region indicated that most landraces were grouped together and had similarity coefficients of over 0.60. It was obvious that genetic relationships among landraces in southwest China had the tendency to associate with their geographic origins.
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2 ij ) /

n , where Hi is the expected het-

erozygosity of the ith allele, and qij refers to the frequency of the jth homozygous allele at ith allele. 6) Wright fix index, defined as inbreeding coefficient F: F = 1-Ho/He. It ranges theoretically from 1 to 1. It is 1 only if landraces are genetically heterozygous. Fit, Fis and Fst are Wright F-statistics parameters [12,13]. Fit and Fis were defined as genetic deviation from Hardy-Weinberg expectation within and among landraces, respectively. Landraces arrive at Hardy-Weinberg equilibrium when Fit and

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Fig. 2

Dendrogram of 54 landraces constructed from SSR marker-based genetic similarity

2.2

Genetic variation of SSR loci within maize landraces A total of 330 alleles were observed at the 42

ples were higher at the same 42 loci. Ae was 3.90 on average and ranged from 2.72 to 7.23. He exhibited a range of variation from 0.56 at 069 to 0.81 at 308707 with the mean of 0.69. Ho averaged 0.37 and ranged from 0.23 (nc133) to 0.65 ( 308707). The most polymorphic loci umc1719 and 308707 revealed a high level of genetic variation.

SSR loci. Allelic diversity at SSR loci varied greatly. The number of alleles A ranged from 3 to 12 with an average of 7.86. In comparison with the bulked DNA samples, the alleles detected in individual DNA samwww.jgenetgenomics.org

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Table 2

The genetic variation of SSR loci and landrace genetic structure

Chromosome 1.02 1.04 1.06 1.10 2.04 2.05 2.08 2.08 3.04 3.05 3.06 3.08 4.00 4.04 4.08 4.11 5.00 5.03 5.03 5.03 5.04 6.00 6.04 6.05 6.07 7.00 7.01 7.02 7.05 7.05 8.02 8.03 8.04 8.06 9.00 9.03 9.05 9.07 10.00 10.02 10.04 10.07 A 3 12 5 10 8 7 7 8 7 8 8 9 8 7 8 8 8 7 7 8 7 7 8 7 7 9 9 9 9 9 10 7 8 7 8 8 6 8 7 9 9 9 7.86 Ae 2.87 5.94 3.71 7.23 4.00 3.09 4.50 3.38 3.82 4.88 3.47 3.22 4.13 3.39 3.61 4.31 4.22 3.71 4.04 3.62 3.65 3.67 4.22 3.30 3.58 3.91 3.44 4.97 2.72 3.50 3.69 3.66 4.47 3.45 2.84 3.84 3.88 4.15 3.61 4.28 4.02 3.63 3.90 He 0.63 0.80 0.67 0.81 0.70 0.63 0.75 0.64 0.70 0.75 0.66 0.63 0.72 0.65 0.67 0.71 0.71 0.67 0.70 0.66 0.69 0.68 0.70 0.65 0.68 0.69 0.66 0.77 0.56 0.65 0.68 0.66 0.74 0.66 0.59 0.70 0.70 0.71 0.67 0.72 0.71 0.67 0.69 Ho 0.32 0.45 0.55 0.65 0.35 0.23 0.48 0.36 0.41 0.37 0.31 0.26 0.38 0.33 0.43 0.29 0.45 0.49 0.32 0.43 0.44 0.39 0.35 0.49 0.41 0.36 0.41 0.39 0.26 0.54 0.34 0.29 0.32 0.34 0.24 0.29 0.27 0.45 0.27 0.38 0.38 0.27 0.37 Fis 0.49 0.44 0.19 0.20 0.51 0.64 0.36 0.44 0.41 0.51 0.53 0.59 0.46 0.49 0.35 0.59 0.38 0.26 0.54 0.35 0.36 0.43 0.50 0.24 0.39 0.77 0.38 0.49 0.54 0.17 0.50 0.56 0.56 0.49 0.59 0.59 0.62 0.37 0.59 0.47 0.46 0.59 0.46 Fit 0.50 0.46 0.25 0.25 0.54 0.67 0.38 0.49 0.45 0.53 0.56 0.63 0.49 0.53 0.40 0.62 0.42 0.32 0.57 0.41 0.39 0.46 0.54 0.29 0.42 0.79 0.42 0.51 0.59 0.24 0.53 0.60 0.58 0.52 0.62 0.61 0.64 0.41 0.62 0.51 0.49 0.62 0.50 Fst 0.03 0.03 0.08 0.06 0.06 0.07 0.04 0.09 0.06 0.05 0.07 0.09 0.06 0.08 0.07 0.07 0.07 0.08 0.07 0.09 0.05 0.07 0.08 0.07 0.06 0.07 0.07 0.04 0.12 0.09 0.07 0.09 0.04 0.07 0.08 0.06 0.06 0.06 0.08 0.06 0.05 0.07 0.07 F 0.50 0.46 0.25 0.25 0.54 0.67 0.38 0.49 0.45 0.53 0.56 0.63 0.49 0.53 0.40 0.62 0.42 0.32 0.57 0.41 0.40 0.46 0.54 0.29 0.42 0.79 0.42 0.51 0.59 0.25 0.53 0.60 0.58 0.52 0.62 0.61 0.64 0.41 0.62 0.50 0.49 0.62 0.50

Locus bnlg1429 umc1719 bnlg1023 phi308707 phi083 nc133 phi090 phi127 phi029 umc1501 phi102228 phi046 phi072 phi096 phi092 phi076 nc130 phi109188 phi008 umc1447 umc1332 phi075 phi031 bnlg1617 phi299852 umc1545 phi057 phi034 phi069 phi051 umc1304 phi115 phi014 umc1161 umc1297 phi065 phi108411 umc2359 phi041 umc1432 phi062 umc1877 Mean

2.3

The genetic structure of maize landraces

Fit was 0.50 on average ranging from 0.24 to 0.79 at corresponding loci. This suggested that an obvious genetic deviation from Hardy-Weinberg expectation occurred among and within landraces. Fst was 0.07 on average, indicating that the among-landrace genetic variation accounted for only 7%, whereas the within-landrace genetic variation for 93%.
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F, Fit, Fis, and Fst were estimated to analyze the genetic structure (Table 2). F varied from 0.25 (bnlg1023) to 0.79 (umc1545), implying that landraces had a typical mixed-mating system and were deficient in heterozygotes. Fis averaged 0.46 ranging from 0.17 ( 051) to 0.77 (umc1545), and

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2.4

Genetic diversity within maize landraces

Discussion

Table 3 summarizes the genetic data with SSRs for each of the 11 maize landraces. A varied from 4.95 to 7.40 and Ae from 2.33 to 4.90. The highest A and Ae were found in DP-11. Similarly, the highest He was in DP-11 (0.82) and the lowest in DP-08 (0.55). Ho was 0.37 within the overall landrace with a small range of variation from 0.33 (DP-66) to 0.39 (DP-68). This indicated that the maximum variation was in DP-11 and the minimum in DP-08. 2.5 Genetic distances between maize landraces

Table 4 shows genetic distances D between the 11 landraces. The genetic distance ranged from the lowest value (0.20) between DP-07 and DP-08 to the highest value (1.26) between DP-03 and DP-59. Consistent with the UPGMA cluster, the mean genetic distance between landraces from different regions was higher thanthat between landraces from the same region.
Table 3 Genetic diversity within 11 maize landraces
Landrace DP-08 DP-11 DP-07 DP-59 DP-40 DP-54 DP-52 DP-66 DP-21 DP-68 DP-03 Overall A 4.95 7.40 5.38 5.50 6.36 6.43 6.40 6.33 6.69 7.07 5.74 7.86 Ae 2.33 4.90 2.89 2.87 3.35 3.14 3.49 3.44 3.64 4.24 3.29 3.90

Among 54 maize landraces for the bulked DNA samples, 6.1 alleles per locus were detected using 42 SSR loci (primers). Liu et al. [14] detected an average of 4.1 alleles with 50 SSR primers in 38 waxy maize landraces. Wu et al.[15] reported an average of 5.4 alleles in popcorn landraces and with 61 SSR primers. Hence, the total number of alleles per locus was higher in this study than previous studies, which suggested a broad genetic base of maize landraces in southwest China. The genetic variability of maize landraces has been affected by various factors throughout their evolutionary history. Outcrossing and fitness-relevant mutations generate intra-population diversity, whereas direct natural or human selection and bottleneck effects lead to an increase in interpopulation diversity [16]. In this study, the withinlandrace genetic variation (93%) was higher than that of the among-landrace (7%). A considerable genetic

He 0.55 0.82 0.66 0.64 0.71 0.69 0.72 0.71 0.74 0.78 0.68 0.69

Ho 0.34 0.39 0.35 0.35 0.39 0.38 0.34 0.33 0.38 0.39 0.35 0.37

Table 4
DP-21 DP-68 DP-40 DP-52 DP-54 DP-03 DP-07 DP-08 DP-59 DP-66

Genetic distances between the 11 landraces

DP-11 0.44 0.21 0.75 0.85 1.03 0.32 0.24 0.25 1.20 1.15 DP-21 0.52 0.53 0.65 0.68 0.65 0.62 0.60 0.75 0.76 DP-68 DP-40 DP-52 DP-54 DP-03 DP-07 DP-08 DP-59

0.74 0.90 1.04 0.26 0.20 0.21 1.22 1.17

0.32 0.36 0.78 0.73 0.73 0.54 0.53

0.32 0.83 0.78 0.78 0.42 0.41

1.12 0.90 0.88 0.29 0.24

0.41 0.45 1.26 1.23

0.20 1.19 1.01

1.10 0.98

0.22

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diversity was revealed within rather than among landraces. Surprisingly, high genetic diversity within-landrace was consistent with the high outcrossing rate of maize. Ranking of landraces based on Ae was identical with their ranking based on He. DP-11 had the highest genetic diversity, with A, Ae, Ho, and He being 7.41, 4.90, 0.82, and 0.39, respectively. In comparison, the lowest values of A, Ae, Ho, and He were observed for DP-08. The landraces with higher genetic diversity DP-11, DP-68, and DP-21 came from Sichuan, while the landraces with lower genetic diversity DP-08 and DP-07 were from Guizhou. The highest genetic diversity occurred in Sichuan, which can be explained by a much longer evolutionary history of maize landraces in Sichuan. This, together with the UPGMA clustering, also supported the hypothesis that maize landraces in southwest China were initially introduced from India into Sichuan [17]. Furthermore, maize landraces in southwest China were distributed from Sichuan to adjacent areas, resulting in the association between genetic variation and geographic origins. The advantages and disadvantages of the bulk DNA sampling analysis have been discussed by Michelmore et al.[18] and Loarce et al [19]. The bulk DNA sampling analysis is labor-saving and rapid but it wipes out the information on individual genotypes, which is needed for estimating the genetic structure and genetic variability within populations. DNA extracted from leaf mixture of 10 individuals was confirmed as a labor saving and efficient approach to analyze genetic relationships among maize populations
[20, 21]

alleles observed in the 11 landraces for the individual DNA sample is higher than that among 54 landraces for bulk DNA samples. Actually, genetic variation harbored in 54 landraces is more than that in 11 landraces. This might be related to the fact that certain SSR loci of bulk DNA could be limited to be detected. Nevertheless, genetic relationships among landraces obtained by the modified Rogers distance are in agreement with those by the UPGMA clustering. This showed that although the bulk DNA sampling analysis wiped genetic diversity among maize landraces partly out, it was effective to evaluate their genetic relationships. The observed and expected heterozygosities within and among landraces showed obvious deviations from Hardy-Weinberg expectation that resulted from heterozygote deficiencies. This finding is expected since farmers are used to renewing maize landraces from year to year with seeds taken from a small number of ears. Consequently, mass selection, which was practiced before development of hybrids, could have led to the deficit of heterozygous individuals. Since inbreeding noticeably affects the length and diameter of the ear, it is likely that farmers unconsciously selected ears from the most heterozygous plants, which prevents genetic drift among landraces and maintains a high level of genetic diversity within landraces. References
1 2 William SK, Michael RC. Essentials of Genetics. Beijing: Higher Education Press, 2002. Darrah LL, Zuber MS. The United States farm corn germplasm base and commercial breeding strategies. Crop Sci, 1986, 26: 11091113. 3 Li Y, Du J, Wang T, Shi Y, Song Y, Jia JJ. Genetic diversity and relationships among Chinese maize inbred lines revealed by SSR markers. Maydica, 2002, 47: 93101. 4 Liu SJ, Rong TZ, Yang JP, Pan GT. Cluster analysis of local maize (Zea mays L.) germplasm in Sichuan based on SSRs. Acta Agronomica Sinica, 2004, 30: 221226 (in Chinese with 5 an English abstract). Yong TZ, Li WS, Yang KC, Zhang B, Zhang SK, Tang HJ,

. Liu et al.[22] studied sampling method

with SSR markers and showed that bulk DNA from 15 individuals could be used to estimate genetic diversity of maize populations. It was also indicated in the preliminary test that bulk DNA from more than 15 individuals resulted in unclear SSR bands, which led to a difficulty in scoring (unpublished data). Therefore, bulk DNA samples of 15 individual plants were used in this study. It was found that the number of

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SSR
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1. 625014 2. 408003 SSR DNA 42 SSR 54 54 256 SSR 2~9 6.1 54 4 54 11 15 165 DNA DNA DNA 42 SSR 11 330 A=7.86 Ae=3.90 He=0.69 H0=0.37F 0.25~0.79 Hardy-Weinberg Fst 0.07 7% 93% SSR

1964 E-mail: yql641@yahoo.com.cn

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