Yukari Akashi

Genetic variation and phylogenetic relationships in East and South Asian melons, Cucumis melo L., based on the analysis of five isozymes

Genetic structure and phylogenetic relationships in East and South Asian melons were analyzed, based on the geographical variation of five isozymes. The analysis of Indian melon accessions showed a continuous variation in seed length, ranging from 4 to 13 mm. Most of the East Asian melons, vars. makuwa and conomon, were classified as the small seed type with seed length shorter than 9 mm. The frequency of the small seed type increased from the west to the east in India. Allelic variation was detected at a total of nine loci of five isozymes among 114 melon accessions. Gene diversity calculated for the nine loci indicated that Indian melon was rich in genetic variation, which decreased from India towards the east. Clear geographical variation was detected in two enzymes, APS and6-PGDH. Pgd-11 and Ap-31 were frequent in India and Myanmar, while most of the melons in Laos, China, Korea and Japan carried Pgd-13 and Ap-33, except var. inodorusin China. Among the latter two alleles, the frequency of Ap-33 was more than 50% in the small seed type in north and east India, indicating that vars. makuwa and conomon were related to the small seed type in these areas. It was also suggested that the small seed type with wet tolerance originated in central India and was selected under wet condition in the east.

Molecular characterization of South and East Asian melon, Cucumis melo L., and the origin of Group Conomon var. makuwa and var. conomon revealed by RAPD analysis

The genetic diversity and relationship among South and East Asian melon Cucumis melo L. were studied by using RAPD analysis of 69 accessions of melon from India, Myanmar, China, Korea, and Japan. The genetic diversity was large in India, and quite small in Group Conomon var. makuwa and var. conomon from East Asia, clearly indicating a decrease in genetic variation from India toward the east. Cluster analysis based on genetic distance classified 17 groups of accessions into two major clusters: cluster I comprising 12 groups of accessions from India and Myanmar and cluster II that included five groups of accessions of Group Conomon var. makuwa and var. conomon from East Asia. Cluster I was further divided into three subclusters, of which subclusters Ib and Ic included small- and large-seed type populations, respectively. Therefore, this division was based on their seed size, not cultivation area. The large-seed type from east India was differently included in the subcluster of small-seed type (Ib). A total of 122 plants of 69 accessions were classified into three major clusters and subclusters: clusters I and II comprised melon accessions mostly from India and Myanmar, and cluster III comprised Group Conomon var. makuwa and var. conomon from East Asia. The frequency of large- and small-seed types was different between clusters I and II, also indicating genetic differentiation between large- and small-seed types. One plant of the small-seed type from east India was differently included in cluster III, and two plants from east India were classified into subcluster IV. These results clearly showed that South Asian melon is genetically differentiated by their seed size, and that small-seed type melon in east India is closely related to Group Conomon var. makuwa and var. conomon.

Phytochrome C Is A Key Factor Controlling Long-Day Flowering in Barley

A red/far-red light photoreceptor controls flowering time in barley by stimulating a flowering promoter gene in the most downstream part of the genetic pathways for flowering. The spring-type near isogenic line (NIL) of the winter-type barley (Hordeum vulgare ssp. vulgare) var. Hayakiso 2 (HK2) was developed by introducing VERNALIZATION-H1 (Vrn-H1) for spring growth habit from the spring-type var. Indo Omugi. Contrary to expectations, the spring-type NIL flowered later than winter-type HK2. This phenotypic difference was controlled by a single gene, which cosegregated only with phytochrome C (HvPhyC) among three candidates around the Vrn-H1 region (Vrn-H1, HvPhyC, and CASEIN KINASE IIα), indicating that HvPhyC was the most likely candidate gene. Compared with the late-flowering allele HvPhyC-l from the NIL, the early-flowering allele HvPhyC-e from HK2 had a single nucleotide polymorphism T1139C in exon 1, which caused a nonsynonymous amino acid substitution of phenylalanine at position 380 by serine in the functionally essential GAF (3′, 5′-cyclic-GMP phosphodiesterase, adenylate cyclase, formate hydrogen lyase activator protein) domain. Functional assay using a rice (Oryza sativa) phyA phyC double mutant line showed that both of the HvPhyC alleles are functional, but HvPhyC-e may have a hyperfunction. Expression analysis using NILs carrying HvPhyC-e and HvPhyC-l (NIL [HvPhyC-e] and NIL [HvPhyC-l], respectively) showed that HvPhyC-e up-regulated only the flowering promoter FLOWERING LOCUS T1 by bypassing the circadian clock genes and flowering integrator CONSTANS1 under a long photoperiod. Consistent with the up-regulation, NIL (HvPhyC-e) flowered earlier than NIL (HvPhyC-l) under long photoperiods. These results implied that HvPhyC is a key factor to control long-day flowering directly.

Molecular analysis of genetic diversity in melon landraces (Cucumis melo L.) from Myanmar and their relationship with melon germplasm from East and South Asia

Genetic diversity of Myanmar melon was evaluated by analysis of 27 RAPD markers and morphological characters using 41 accessions of melon landraces of which 36 accessions were small-seed type. The gene diversity was 0.239, higher than for group Conomon from East Asia and equivalent to Indian melon populations. Melon accessions were classified into six major clusters. The largest cluster IV comprised mainly group Conomon which was closely related to cluster V consisting of mainly group Agrestis. Most of the accessions of group Cantalupensis were grouped into clusters II or VII and were distantly related to groups Conomon and Agrestis. The genetic relationship to melon accessions from neighboring countries was analyzed. The 24 accessions of clusters IV and V were mostly clustered together with small-seed type melon of India, but the 14 accessions of clusters VI and VII were mostly clustered together with large-seed type melon of India. These results indicated that the genetic diversity of Indian melon is conserved in Myanmar. Genetic introgression among melon groups through spontaneous hybridization was also indicated and was considered important to maintain or increase the genetic diversity in Myanmar.

Diversification and genetic differentiation of cultivated melon inferred from sequence polymorphism in the chloroplast genome.

Molecular analysis encouraged discovery of genetic diversity and relationships of cultivated melon (Cucumis melo L.). We sequenced nine inter- and intra-genic regions of the chloroplast genome, about 5500 bp, using 60 melon accessions and six reference accessions of wild species of Cucumis to show intra-specific variation of the chloroplast genome. Sequence polymorphisms were detected among melon accessions and other Cucumis species, indicating intra-specific diversification of the chloroplast genome. Melon accessions were classified into three subclusters by cytoplasm type and then into 12 subgroups. Geographical origin and seed size also differed between the three subclusters. Subcluster Ia contained small-seed melon from Southern Africa and South and East Asia and subcluster Ib mainly consisted of large-seed melon from northern Africa, Europe and USA. Melon accessions of subcluster Ic were only found in West, Central and Southern Africa. Our results indicated that European melon groups and Asian melon groups diversified independently and shared the same maternal lineage with northern African large-seed melon and Southern African small-seed melon, respectively. Cultivated melon of subcluster Ic may have been domesticated independently in Africa. The presence of 11 cytoplasm types in Africa strongly supported African origin of cultivated melon and indicated the importance of germplasm from Africa.