Assessment of genetic relationships among Cucumis spp. by SSR and RAPD marker analysis

The first successful production of a sterile interspecific hybrid obtained from a cross between Cucumis hystrix Chakr. (2n = 2x = 24) and Cucumis sativus var. sativus L. (2n =2x = 14), and its subsequent fertility restoration through chromosome doubling provide an effective means for investigating genetic relationships among Cucumis spp. In this study, random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to investigate relationships among C. s. var. sativus L., C. s. var. hardwickii (R.) Alef., C hystrix, C. hytivus Chen & Kirkbride (the amphidiploid species from chromosome doubling of the C. sativus x C. hystrix interspecific hybrid, 2n = 38), C. melo (2n =2x = 24) and C. metuliferus Meyer and Naudin (2n =2x= 24). A total of 109 SSR bands and 398 RAPD primed sites were used to calculate Jaccard's distance coefficients for cluster analysis using a unweighted pair‐group method using an arithmetic averaging (UPGMA) algorithm. The genetic relationships identified using SSR and RAPD markers were highly concordant, such that the correlation between SSR and RAPD genetic distance (GD) estimates was r = 0.94. SSR and RAPD analysis of 22 accessions allowed for their grouping into two distinct groups designated as CS and CM. While group CS consisted of 11 C. sativus genotypes, and the C. hytivus and C. hystrix accessions, group CM included six C. melo genotypes and C. metuliferus. The GD values between C. hystrix and C. sativus ascribed by SSR and RAPD matrices were 0.59 and 0.57, respectively. These GDs were smaller than those detected between C. hystrix and C melo (0.87 and 0.70 derived from SSR and RAPD markers, respectively).     

Heterosis performance of yield and fibre quality in F1 and F2 hybrids in upland cotton

Because of the difficulty of producing F₁ hybrid seeds by hand emasculation and pollination, wide use of heterosis in cotton production has been limited in China. The objective of this study was to evaluate the potential of F₂ hybrids for yield and fibre quality. A half diallel involving eight parents and their F₁ and F₂ hybrids was grown in replicated studies at Linqing and Nanjing in 1999 and Nanjing in 2000. Yield and fibre quality was determined for all 64 entries. Fibre quality was also determined for parents and F₁s, but only for Zhongmiansuo 28 (ZMS28), Xiangzamian 2 (XZM2) and Wanmian 13 (WM13) F₂s. These three F₂ hybrids are extensively planted in China and provide experimental controls with which to compare the performance of new hybrids. Average yield heterosis for F₁s and F₂s was 15.9 and 9.2%, respectively. Inbreeding depression for yield varied but some F₂s greatly out‐yielded the best variety. Average F₁ heterosis was 6.7, 6.2 and 2.9%, respectively for number of bolls per unit area, boll weight, and lint percentage. The average F₂ heterosis for the same traits was 4.4, 3.3 and 1.6%, respectively. F₁ heterosis for fibre traits was low. In general, parental average was a good indicator of the yield and fibre quality of F₁ hybrids. These encouraging results suggest there is sufficient heterosis for yield to use F₂s in China.     

RFLP/AFLP mapping of a brown planthopper (Nilaparvata lugens Stål) resistance gene Bph1 in rice

We have constructed a linkage map of the rice brown planthopper (BPH)resistance gene, Bph1. RFLP and AFLP markers were selected by thebulked segregant analysis and used in the mapping study of 262 F₂sthat were derived from a cross of `Tsukushibare', a susceptible japonica cultivar, and `Norin-PL3', an authentic japonicaBph1-introgression line. Twenty markers were mapped within a 28.9-cMregion containing the Bph1 locus on the long arm of rice chromosome12. Combining the result of segregation analysis of BPH resistance by themass seedling test and that of the markers, the Bph1 locus wasmapped within a 5.8-cM region between two flanking markers. The closestAFLP markers, em5814N and em2802N, was at 2.7 cM proximal to theBph1 locus. Together with the previously constructed high-resolutionmap of bph2 locating the locus at ca. 10 cM proximal to the Bph1 locus, this improved version of the linkage map would facilitatepyramiding these two important BPH resistance genes.     

Production and characterization of Raphanus sativus-Brassica rapa monosomic chromosome addition lines

Breeding of Raphanus sativus‐Brassica rapa monosomic chromosome addition lines (MALs, 2n = 19) was carried out by backcrossing the synthesized amphidiploid line, Raphanobrassica (R. sativus×B. rapa, 2n = 38, RRAA, line RA89) with R. sativus cv. ‘Shogoin’ (2n = 18, RR). In the first cross of Raphanobrassica× radish, four sesquidiploidal BC₁ plants (2n = 28, RRA, RA89‐36‐1, RA89‐31‐1, RA89‐31‐2, RA89‐31‐3) were successfully developed. In these plants, the chromosome configurations of 9II + 10I and 10II + 8I were observed frequently at first metaphase (MI) of meiosis in pollen mother cells (PMCs). The RA 89‐36‐1 plant produced many seeds in the reciprocal backcrosses with radish. About 50% of the BC₂ plants obtained from the cross of RA89‐36‐1 plant × radish were 2n = 19 plants, followed by 2n = 18 plants (24%) and 2n = 20 plants (19%). In the reciprocal cross, 2n = 19 plants were also developed at the rate of 40%. From analysis of specific morphological traits, 2n = 19 plants were classified into eight types (a‐h). When 25 selected primers were used in polyacrylamide gel electrophoresis, random amplified polymorphic DNA (RAPD) markers derived from B. rapa for each type of MAL were detected in numbers between three for e‐type and 16 for b‐type. RAPD markers specific for each type alone were from one (OPE 05‐344) for h‐type to nine for b‐type. In the g‐type, no marker specific to this type alone was observed. However, 19 bands were common between at least two types. These MAL plants exhibited predominantly the chromosome configuration of 9II + 1I at MI of PMCs, pollen and seed fertility being the same level as the radish cv. ‘Shogoin’. From the morphological traits and DNA markers, eight different MAL types among 10 expected were identified.     

Differentiation of the high molecular weight glutenin subunit D t x 2.1 of Aegilops tauschii indicated by partial sequences of its encoding gene and SSR markers

The devastating late blight pathogen Phytophthora infestans infects foliage as well as tubers of potato. To date, resistance breeding has often focused on foliage blight resistance, but tuber blight resistance is becoming more and more important in cultivated potatoes. In this study, a reliable tuber assay for resistance assessment was developed and foliage and tuber blight resistance (R) was compared in four mapping populations. In the RH4X-103 population, tuber blight resistance inherited independently from foliage blight resistance. Three specific R genes against P. infestans were segregating. The Rpi-abpt and R3a genes function as foliage-specific R genes, whereas the R1 gene acts on both foliage and tuber. In the segregating populations SHRH and RH94-076, tuber and foliage blight resistance correlated significantly, which suggests that resistance in foliage and tuber is conferred by the same gene (could be R3b) and quantitative trait loci (QTL), respectively. In the CE population neither tuber nor foliage resistance was observed.     

QTL analysis of cooked rice grain elongation, volume expansion, and water absorption using a recombinant inbred population

The traits of elongation, volume expansion, and water absorption are very important in determining the quality of cooked rice grains. In this study, quantitative trait loci (QTL) analysis of these traits was performed using a recombinant inbred population derived from a cross between two indica cultivars, ‘Zhenshan 97’ and ‘Minghui 63 ,’ which are the parents of the most widely grown hybrid rice in China. Using a linkage map based on 221 molecular marker loci covering a total of 1796 cM, a total of 33 QTLs were identified for the nine traits tested. QTLs were detected on chromosomes 1– 3 , 5– 9 , and 11 , respectively. The QTLs identified included three for cooked rice grain length elongation (chromosomes 2 , 6 , and 11), six for width expansion (chromosomes 1‐ 3 , 6 , 9 , and 11) and two for water absorption (chromosomes 2 and 6). Interestingly, a single QTL located near the wx gene on chromosome 6 seemed to influence all the traits tested for the cooked rice quality.     

Discovery of new male-sterile cytoplasm sources and development of a new cytoplasmic-nuclear male-sterile line NJCMS3A in soybean

Most of the cytoplasmic-nuclear male-sterile (CMS) lines of soybean were developed only from a limited cytoplasm sources and performed not as good as required in hybrid seed production, therefore, to explore new male-sterile cytoplasm sources should be one of the effective ways to improve the pollination and hybridization for a better pod-set in utilization of heterosis of soybeans. In the present study, total 80 crosses between 70 cultivated and annual wild soybean accessions and three maintainers (N2899, N21249, and N23998) of NJCMS1A were made for detecting potential new sources with male-sterile cytoplasm. The results showed that in addition to the crosses with N8855.1 (the cytoplasm donor parent of NJCMS1A) and its derived line NG99-893 as cytoplasm parent, there appeared three crosses, including N21566 × N21249 and N23168 × N21249, with male-sterile plants in their progenies. According to the male fertility performance of backcrosses and reciprocal crosses with the tester N21249, the landrace N21566 and annual wild soybean accession N23168 were further confirmed to have male-sterile cytoplasm. Accordingly, it was understood that the source with male-sterile cytoplasm in soybean gene pool might be not occasional. The results also showed that the genetic system of male sterility of the newly found cytoplasm source N21566 was different from the old cytoplasm source N8855.1, while N23168 was to be further studied. Based on the above results, the derived male-sterile plants from [(N21566 × N21249) F₁ × N21249] BC₁F₁ were back-crossed with the recurrent parent N21249 for five successive times, and a new CMS line and its maintainer line, designated as NJCMS3A and NJCMS3B, respectively, were obtained. NJCMS3A had normal female fertility and stable male sterility. Its microspore abortion was mainly at middle uninucleate stage, earlier than that of NJCMS1A and NJCMS2A. The male fertility of F₁s between NJCMS3A and 20 pollen parents showed that 7 accessions could restore its male fertility and other 13 could maintain its male sterility. The male sterility of NJCMS3A and its restoration were controlled by one pair of gametophyte male-sterile gene according to male fertility segregation of crosses between NJCMS3A and three restorers. The nuclear gene(s) of male sterility in NJCMS3A appeared different from the previously reported CMS lines, NJCMS1A and NJCMS2A. The development of NJCMS3A demonstrated the feasibility to discover new CMS system through choosing maintainers with suitable nuclear background.     

Search for partial resistance against Puccinia hordei in barley landraces from the Fertile Crescent

A collection of 111 barley landraces from the Fertile Crescent was screened for resistance to barley leaf rust in the field and under controlled conditions. Large variation was observed for disease severity under field conditions. Accessions with high resistance because of hypersensitivity were identified. Also segregation was observed in some accessions, with individual plants showing hypersensitive reactions (IT ≤ 6). Partial resistance due to a reduction of infection in spite of a compatible infection was commonly found (19%). Resistance of 12 accessions selected for their low disease severity and high IT, was shown to be due to a prolonged latency period and increased percentage of early aborted colonies not associated with host cell necrosis. A high correlation was observed between the microscopic and macroscopic components of partial resistance.     

Two chromosome segments confer multiple potyvirus resistance in maize

Potyviruses cause serious yield losses in maize production worldwide. While the maize dwarf mosaic virus (MDMV) predominates in the USA, sugarcane mosaic virus (SCMV) is a major pathogen in China and Germany. In previous studies, inbred FAP1360A revealed complete resistance against both MDMV and SCMV. Two major SCMV resistance genes, Scmv1 and Scmv2, were located on chromosomes 6 and 3, respectively, in populations derived from crosses with the susceptible inbred line F7. For validation of these results obtained in segregating backcross‐ or F_2:3‐populations, near‐isogenic lines to F7 have been produced after one initial cross to FAP1360A by repeated backcrossing to F7, phenotypic selection for SCMV resistance, and marker‐assisted selection for the Scmv1 and Scmv2 regions from FAP1360A. The near‐isogenic line F7R has been studied in detail both at the genomic level and for resistance to different potyviruses. Based on 112 polymorphic simple sequence repeat markers, F7R received genomic segments introgressed from FAP1360A exclusively in the Scmv1 and Scmv2 chromosomal regions. F7R conferred complete resistance to SCMV and MDMV, but also to zea mosaic virus and to systemic infection by wheat streak mosaic virus. FAP1360A, F7, F7R were not systemically infected by high plains virus. Thus, introgression of Scmv1 and Scmv2 from FAP1360A into F7 was sufficient to generate the first potyvirus multiresistant European Flint line reported so far.     

Pyramiding of Xa7 and Xa21 for the improvement of disease resistance to bacterial blight in hybrid rice

‘Minghui 63’ is a restorer line widely used in hybrid rice production in China for the last two decades. This line and its derived hybrids, including ‘Shanyou 63’, are susceptible to bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (Xoo). To improve the bacterial blight resistance of hybrid rice, two resistance genes Xa21 and Xa7, have been introgressed into ‘Minghui 63’ by marker‐assisted selection and conventional backcrossing, respectively. The single resistance gene‐introgressed lines, Minghui 63 (Xa21) and Minghui 63 (Xa7) had higher levels of resistance to bacterial blight than their derived hybrids, Shanyou 63 (Xa21) or Shanyou 63 (Xa7). Both Xa21 and Xa7 showed incomplete dominance in the heterozygous background of rice hybrids by infection with GX325 and KS‐1‐21. The improved restorer lines, with the homozygous genotypes, Xa21Xa21 or Xa7Xa7, were more resistant than their hybrids with the heterozygous genotypes Xa21xa21 or Xa7xa7. To further enhance the bacterial blight resistance of ‘Minghui 63’ and its hybrids, Xa21 and Xa7 were pyramided into the same background using molecular marker‐aided selection. The restorer lines developed with the resistance genes Xa21 and Xa7, and their derived hybrids were evaluated for resistance after inoculation with 10 isolates of pathogens from China, Japan and the Philippines, and showed a higher level of resistance to BB than the restorer lines and derived hybrids having only one of the resistance genes. The pyramided double resistance lines and their derived hybrids have the same high level of resistance to BB. These results clearly indicate that pyramiding of dominant genes is a useful approach for improving BB resistance in hybrid rice.