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Molecular Analysis of the Genetic Diversity of Chinese Hami Melon and Its Relationship to the Melon Germplasm from Central and South Asia

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Molecular Analysis of the Genetic Diversity of Chinese Hami Melon and Its Relationship to the Melon Germplasm from Central and South Asia J. Japan. Soc. Hort. Sci. 80 (1): 52–65. 2011. Available online at www.jstage.jst.go.jp/browse/jjshs1 JSHS © 2011 Molecular Analysis of the Genetic Diversity of Chinese Hami Melon and Its Relationship to the Melon Germplasm from Central and South Asia Yasheng Aierken1,2, Yukari Akashi1, Phan Thi Phuong Nhi1, Yikeremu Halidan1, Katsunori Tanaka3, Bo Long4, Hidetaka Nishida1, Chunlin Long4, Min Zhu Wu2 and Kenji Kato1* 1Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan 2Hami Melon Research Center, Xinjiang Academy of Agricultural Science, Urumuqi 830000, China 3Research Institute for Humanity and Nature, Kyoto 603-8047, Japan 4Kunming Institute of Botany, CAS, Heilongtan, Kunming, Yunnan 650204, China Chinese Hami melon consists of the varieties cassaba, chandalak, ameri, and zard. To show their genetic diversity, 120 melon accessions, including 24 accessions of Hami melon, were analyzed using molecular markers of nuclear and cytoplasmic genomes. All Hami melon accessions were classified as the large-seed type with seed length longer than 9 mm, like US and Spanish Inodorus melon. Conomon accessions grown in east China were all the small- seed type. Both large- and small-seed types were in landraces from Iran, Afghanistan, Pakistan, and Central Asia. Analysis of an SNP in the PS-ID region (Rpl16-Rpl14) and size polymorphism of ccSSR7 showed that the melon accessions consisted of three chloroplast genome types, that is, maternal lineages. Hami melon accessions were T/338 bp type, which differed from Spanish melon and US Honey Dew (T/333 bp type), indicating a different maternal lineage within group Inodorus. The gene diversity (D), calculated from random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) polymorphism, was 0.476 in 120 melon accessions; the largest diversity was in Central Asian accessions (D = 0.377) but was low for Hami melon accessions (D = 0.243), even though Hami melon has diverse morphological traits, earliness, and shelf life. Reflecting such small genetic diversity, Hami melon accessions of vars. ameri and zard were grouped into cluster II, except for one accession, by the unweighted pair group method and the arithmetic mean (UPGMA) cluster analysis. Variety chandalak with distinct characters, such as early maturing and poor shelf life, was assigned to clusters IV and VI, indicating inter-varietal genetic differentiation within Hami melon. Three accessions from Turkmenistan and Afghanistan, with large seeds and T/338 bp type of chloroplast genome, were classified as cluster II with Hami melon accessions of vars. ameri and zard. We therefore concluded that Hami melon may have been transmitted from the west. The small-seed type melon of group Conomon grown in east China may have been introduced into China independently of Hami melon, because it had the A/338 bp type of the chloroplast genome and was clustered distantly from Hami melon according to nuclear genome analysis. Key Words: chloroplast genome, Cucumis melo, genetic diversity, Hami melon, SSR. with a total production of 9.7 million tons a year. Chinese Introduction melon is generally divided into two types based on the Melon (Cucumis melo L.) is one of the most important thickness of the fruit skin: thick-skinned melon is horticultural crops in China, and is cultivated in 353 kha classified as Group Inodorus or Cantalupensis among seven horticultural groups defined by Munger and Robinson (1991), and thin-skinned melon is classified Received; July 5, 2010. Accepted; August 12, 2010. as Group Conomon. These two types also differ in This study was partly supported by a Grant-in-Aid for International cultivation area. Thick-skinned melon is mostly Scientific Research of Ministry of Education, Science, Culture and produced in Xinjiang (Xinjiang Uyghur Autonomous Sports, Japan (No. 19255009), and JSPS Asian CORE Program. This is Contribution number 23 from the Sato Project of Research Institute Region), and thin-skinned melon in eastern China. Both for Humanity and Nature (RIHN), Japan. types are cultivated between these two areas in Qinghai, * Corresponding author (E-mail: [email protected]). Gansu, and Shaanxi provinces. 52 J. Japan. Soc. Hort. Sci. 80 (1): 52–65. 2011. 53 Xinjiang is in north-west China, and is one-sixth of var. chandalak is consumed in Xinjiang and is not China’s national land. It has a dry continental climate exported because this type is unsuitable for long-distance with great extremes of winter and summer temperature. transportation because of its short shelf life. However, Rainfall is scant, and the annual precipitation is less than var. zard and the mid-season maturing group of var. 500 mm in north Xinjiang and less than 100 mm in south ameri have a long (> six months) to moderately long Xinjiang. It is below 40 mm in Tulufan and Hami, famous (one month) shelf life and are exported as Hami melon for producing Hami melon, in the eastern part of to foreign countries, as well as to other provinces of Xinjiang. The daily fluctuation of air temperature is China. Taking such diversification into account, Pitrat large, and the average difference between day and night et al. (2000) proposed to classify varieties cassaba and temperature is from 11.3°C to 14.8°C depending on the zard as var. inodorus (group Inodorus), and the varieties area. Other important features of the climate are strong chandalak and ameri as independent varieties. However, sunshine and long sunshine duration. These conditions the classification was primarily based on morphological are advantageous for producing sweet melon. Thick- and physiological characters, and the genetic diversity skinned melon called Hami melon (“Hami Gua” in within each variety, and the genetic relationship between Chinese) is cultivated in 41 kha with a total production varieties, has remained unclear. of 1.1 million tons a year. In the last decade, the genetic diversity and Local landraces of Hami melon are rich in diversity, phylogenetic relationship between various groups of and 101 local landraces of melon were collected in cultivated melon have been studied using analysis of Xinjiang to study melon germplasm in the 1950s and molecular polymorphism, which can be easily detected 1980s (Wu, 1982). These landraces were classified as using various types of DNA markers, such as random var. cassaba Pang Greb (round fruits with three or five amplified polymorphic DNA (RAPD: Dhillon et al., carpels, one accession), var. chandalak Pang Greb (extra 2007; López-Sesé et al., 2003; Luan et al., 2008; Nakata early Guadan melon, five accessions), var. ameri Pang et al., 2005; Soltani et al., 2010; Staub et al., 2000, 2004; Greb (summer melon, 70 accessions), and var. zard Pang Stepansky et al., 1999; Tanaka et al., 2007; Yi et al., Greb (winter melon, 25 accessions) (Lin, 1991). Figure 1 2009), simple sequence repeats (SSRs: Dhillon et al., shows photographs of typical fruits of these varieties. 2007; Garcia-Mas et al., 2004; López-Sesé et al., 2002; Var. ameri consists of early maturing and mid-season Monforte et al., 2003; Nakata et al., 2005; Staub et al., maturing groups. Similarly, var. zard includes autumn 2000), and amplified fragment length polymorphism melon and late maturing winter melon groups. The (AFLP: Garcia-Mas et al., 2000; Yashiro et al., 2005). sowing time is from the end of March (south Xinjiang) These results showed distinct genetic differentiation to the middle of April (north Xinjiang). The harvest of between subsp. agrestis and subsp. melo, which were var. chandalak starts from 10 June, and the latest defined by Pitrat et al. (2000) and Jeffrey (2001): subsp. maturing winter melon (var. zard) is harvested in agrestis has short hairs on the ovary and subsp. melo October; the fruits of var. zard are stored until May of has long hairs on the ovary. Seed size is also an important the next year. Cultivation of these types facilitates the character to distinguish these two subspecies. Small-seed year-round supply of Hami melon. Among these types, type melon groups Conomon and Agrestis with seed length shorter than 9 mm were classified as subsp. agrestis, and large-seed type melon groups Catalupensis, Inodorus and Flexuosus with seed length over 9 mm were classified as subsp. melo (Akashi et al., 2002). However, the genetic differentiation between varieties is not clear within each subspecies, and subsp. melo accessions of groups Catalupensis, Inodorus and Flexuosus have been clustered together (López-Sesé et al., 2003; Soltani et al., 2010; Stepansky et al., 1999). In contrast, the geographical variation in each horticultural group is often significant, as indicated by López-Sesé et al. (2003) who detected clear differences between Spanish landraces and US-EU improved cultivars of Inodorus, although most Inodorus accessions analyzed in these studies were selected from Europe and USA, and little attention was paid to accessions from Central Asia and China. Although Monforte et al. (2003) analyzed one accession each of varieties chandalak and ameri, these accessions were from Russia and Uzbekistan, respectively. Therefore, little is known about Fig. 1. Photographs of fruits of four varieties of Hami melon. the genetic diversity and genetic structure of Hami 54 Y. Aierken, Y. Akashi, P.T.P. Nhi, Y. Halidan, K. Tanaka, B. Long, H. Nishida, C. Long, M.Z. Wu and K. Kato melon, irrespective of its importance as a novel genetic included as Central Asian accessions, Spanish acces- resource for a wide range of shelf life, flavor, high sugar sions, US Inodorus, and US Cantalupensis, respectively, content, large fruit, etc. (Lin, 1991; Liu et al., 2004). in this study, even though Tanaka et al. (2007) used them In Central Asian countries located to the west of as reference accessions. Xinjiang, group Inodorus is mainly grown, and group For DNA analysis, total DNA was extracted from one Conomon vars.
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