No abstract

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 IIa), 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 (39, 59-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.

Natural variation in wheat requirement of long exposures to cold temperatures to accelerate Xowering (vernalization) is mainly controlled by the Vrn-1, Vrn-2, Vrn-3, and Vrn-4 loci. The Wrst three loci have been well characterized, but limited information is available for Vrn-4. So far, natural variation for Vrn-4 has been detected only in the D genome (Vrn-D4), and genetic stocks for this gene are available in Triple Dirk (TDF, hereafter). We detected heterogeneity in the Vrn-1 alleles present in diVerent TDF stocks, which may explain inconsistencies among previous studies. A correct TDF seed stock from Japan carrying recessive vrn-A1, vrn-B1, and vrn-D1 alleles was crossed with three diVerent winter cultivars to generate F 2 mapping populations. Most of the variation in Xowering time in these three populations was controlled by a single locus, Vrn-D4, which was mapped within a 1.8 cM interval Xanked by markers Xcfd78 and Xbarc205 in the centromeric region of chromosome 5D. A factorial ANOVA for heading time using Vrn-D4 alleles and vernalization as factors showed a signiWcant interaction (P < 0.0001), which conWrmed that the Vrn-D4 eVect on Xowering time is modulated by vernalization. Comparison of the diVerent Triple Dirk stocks revealed that Vrn-B1, Vrn-D1, and Vrn-D4 all have a small residual response to vernalization, but Vrn-D4 diVers from the other two in its response to short vernalization periods. The precise mapping and characterization of Vrn-D4 presented here represent a Wrst step toward the positional cloning of this gene.

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 smallseed 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.

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 smallseed 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.