Yoshiteru Sakata

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.

Chloroplast DNA analysis of eggplant (Solanum melongena) and related species for their taxonomic affinity

SummaryTotal chloroplast DNA (cpDNA) from Solanum incanum, a wild relative of eggplant, was used to probe total DNA of Solanum melongena (eggplant). The DNA fragments detected were the same as observed using purified chloroplast DNA. Chloroplast DNAs were also analysed for nine species of Solanum that are cross-compatible with eggplant: S. aethiopicum, S. anguivi, S. gilo, S. incanum, S. indicum, S. integrifolium, S. macrocarpon, S. olivare and S. panduriforme.Restriction fragments generated by eight enzymes were recorded as present or absent, and a matrix for all fragment positions, species and enzymes was used for cluster analysis. In the resulting dendrogram, the species tested formed three distinct groups: (1) S. aethiopicum, S. anguivi, S. gilo, S. indicum, S. integrifolium and S. olivare, (2) S. incanum, S. melongena and S. panduriforme, (3) S. macrocarpon. Six species of the first group belonging to section Oliganthes appears more closely related to the second group members belonging to section Melongena than does S. macrocarpon, which also belongs to section Melongena. Within the second group, S. panduriforme is slightly more like eggplant than is S. incanum.

Chloroplast DNA diversity in brinjal eggplant (Solanum melongena L.) and related species

Chloroplast DNA (cpDNA) samples of brinjal eggplant (S. melongena) and representative related species including S. incanum sensu lato (or S. campylacanthum sensu stricto), S. lichtensteinii, S. marginatum, S. macrocarpon, S. anguivi and S. aethiopicum and also S. nigrum as an outgroup taxon, were digested by 14 restriction enzymes and analyzed by using electrophoresis and a cpDNA probe. All the species used here were clearly separated in the cpDNA analysis, except the pair S. anguivi and S. aethiopicum. From the dendrogram constructed by the unweighted pair-group method, it is suggested that S. incanum is the closest to S. melongena and the next closest species is S. macrocarpon followed by S. aethiopicum (and S. anguivi), S. lichtensteinii, S. marginatum and finally the outgroup taxon S. nigrum. The tree derived by the neighbour-joining method suggests phyletic relationships that agree with those indicated by crossability and seed coat anatomy, but conflict with conventional classifications based on morphology. In particular, members of sections Oliganthes and Melongena are not separated and no cpDNA variation was found within either of the morphologically diverse cultigens, S. aethiopicum and S. melongena. Paradoxically, the morphologically similar species S. incanum, S. lichtensteinii and S. marginatum have diverged greatly in their cpDNA. The significance of these results is discussed.

Identification of quantitative trait loci for downy mildew resistance in cucumber (Cucumis sativus L.)

Downy mildew, caused by Pseudoperonospora cubensis (Berk. & Curt.) Rostov, is one of the most economically important foliar diseases in cucumber (Cucumis sativus L.). Cucumber line CS-PMR1, derived from self-pollination of USDA Plant Introduction 197088, has a high level of resistance to downy mildew and is considered to be promising breeding material. In this study, we performed quantitative trait locus (QTL) analysis for downy mildew resistance using 111 recombinant inbred lines (RILs) derived from a cross between CS-PMR1 and the old Japanese cultivar Santou, which exhibits moderate resistance. The resistance of the RILs and their parents was evaluated by diverse methods using different plant organs (cotyledons, true leaves), stages (seedlings and adult plants), and evaluation criteria (lesion expansion and extent of sporulation). The high resistance of CS-PMR1 was associated with many QTLs with relatively small effects, whereas the moderate resistance of Santou was associated with one major QTL and possibly two others with relatively small effects. In all assays, the major QTL at which the Santou allele was associated with increased resistance had the largest effect. This QTL allele from Santou and several of the most effective QTL alleles identified in CS-PMR1 should be highest priority for selection to efficiently breed new cultivars that carry adequate levels of downy mildew resistance.

Chloroplast DNA diversity in eggplant (Solanum melongena) and its related species S. incanum and S. marginatum

SummaryTotal chloroplast DNA (cpDNA) from Solanum incanum, a wild relative of eggplant, was used to probe total DNAs from 27 accessions of S. melongena (eggplant), S. incanum sensu lato and S. marginatum, all of which are quite similar in their morphology.There is a wide diversity in plastotypes within S. incanum sensu lato. On the other hand, only one restriction fragment pattern difference was detected between S. melongena and S. incanum sensu lato.The restriction fragment patterns generated by eight enzymes were recorded as present or absent, and a matrix for all fragment positions, enzymes and accessions was used for cluster analysis. In the dendrogram, it is suggested that S. marginatum is not nearly as closely related to S. melongena and S. incanum sensu lato as previously supposed, and some of accessions treated as S. incanum sensu lato, originated from Southern Africa, should be called S. lichtensteinii.

Resistance in melon to Cucurbit chlorotic yellows virus, a whitefly-transmitted crinivirus

Fifty-one melon (Cucumis melo) accessions that originated from India, Pakistan and Bangladesh were evaluated for resistance to Cucurbit chlorotic yellows virus (CCYV), a newly emerged species in the genus Crinivirus. CCYV was inoculated using sweet potato whitefly (Bemisia tabaci) biotype Q. Accessions, JP 138332, JP 216154, JP 216155, JP 216751 and JP 91204, showed no or faint symptoms, although CCYV was detected from the non-inoculated upper leaves by reverse transcription-polymerase chain reaction (RT-PCR). The five accessions were subjected to quantitative RT-PCR to analyze relative accumulation of CCYV RNA. All accessions except JP 138332 had levels of CCYV RNA accumulation comparable to the commercial variety, ‘Earl’s Seine’, which was used as a control. JP 138332 showed a much lower CCYV RNA accumulation. Numbers of B. tabaci biotype Q on JP 138332 did not differ from ‘Earl’s Seine’, in antixenosis tests, and the result suggested the resistance to CCYV was not due to antixenosis. Consequently, five accessions are of interest for development of resistant varieties. In particular, JP 138332 possesses a promising resistant trait for CCYV, which might be associated with inhibition of virus multiplication.

Effect of temperature on symptom expression and viral spread of Melon yellow spot virus in resistant cucumber accessions

Melon yellow spot virus (MYSV), a member of the genus Tospovirus, is a devastating thrips-transmitted virus of cucurbits in Japan. Recently, we reported that cucumber accessions originating from South Asia, in particular Southeast Asia, had moderate resistance to MYSV. Here, we investigated the effect of three temperatures (20°C, 25°C, and 30°C) on symptom expression and viral spread of MYSV in plants of resistant cucumber accessions. No systemic infection developed in resistant cucumber plants after inoculation with melon isolate MYSV-S at low temperature (20°C); viral spread of MYSV-S and cucumber isolate MYSV-FuCu05P in inoculated cotyledons was suppressed. In contrast, higher incubation temperatures (25°C and 30°C) facilitated viral spread in inoculated cotyledons and systemic infection of MYSV-S. These data suggest that the resistance to MYSV of resistant cucumber accessions is temperature dependent.