Genetic analysis and identification of two soybean mosaic virus resistance genes in soybean [Glycine max (L.) Merr]

Soybean mosaic virus (SMV) commonly affects soybean production worldwide, and the SC18 strain has been widespread in China. This study aimed to characterize and map the SC18 resistance genes present in soybean cultivars ‘Kefeng No. 1’ and ‘Qihuang 22’. Inheritance analysis revealed that two independent single dominant genes in Kefeng No. 1 and Qihuang 22 confer resistance to SC18. Using simple sequence repeat (SSR) markers and bulked segregant analysis, the Kefeng No. 1 and Qihuang 22 resistance genes were located on soybean chromosomes 2 and 13, respectively. We further screened two populations of recombinant inbred lines with 32 SSR markers in the target region, where the resistance gene in Kefeng No. 1 was fine mapped to an 80‐kb region containing six putative genes. Sequence and expression analyses of these genes revealed that SMV resistance in Kefeng No. 1 was probably attributable to three of the candidate genes (i.e. Glyma.02G127800, Glyma.02G128200 and Glyma.02G128300). Collectively, the results of this study will greatly facilitate the cloning of SC18 resistance genes and marker‐assisted breeding of SMV‐resistant soybean cultivars.     

Identification and characterization of a RAPD/SCAR marker linked to a resistance gene for soybean mosaic virus in soybean

Soybean line `ICGR95-5383' [Glycinemax (L.) Merr.] is a newly releasedgermplasm from China and is resistant (R)to soybean mosaic virus (SMV). ICGR95-5383was crossed to the susceptible (S)cultivars `HB1', `Tiefeng21', `Amsoy', and`Williams' to investigate the inheritanceof SMV resistance. The F<sub>1</sub> and F<sub>2</sub>plants were inoculated with SMV-3 (the mostvirulent) strain from Northeast China. Theresults showed that F<sub>1</sub> plants from thefour R × S crosses were necrotic (N) andall F<sub>2</sub> populations segregated in a3(R+N):1S ratio, indicating thatICGR95-5383 carries a single gene withincomplete dominance for resistance to SMV. In a bulked segregant analysis (BSA) of theF<sub>2</sub>population from ICGR95-5383 × HB1, a codominant RAPD marker,OPN11<sub>980/1070</sub>, was found to be linkedto the resistance gene in ICGR95-5383. The980-base pair (bp) fragment OPN11<sub>980</sub>was amplified in the R parent ICGR95-5383,R bulk, and resistant F<sub>2</sub> plants. Theother 1070-bp fragment OPN11<sub>1070</sub> wasamplified in the S parent HB1, S bulk, andsusceptible F<sub>2</sub>plants.OPN11<sub>980/1070</sub> was amplified in theF<sub>1</sub> plants and the necroticF<sub>2</sub> plants from the R×S cross.Segregation analysis of the RAPD marker inthe F<sub>2</sub> population revealed that themarker OPN11<sub>980/1070</sub> is closely linkedto the resistance gene with a map distanceof 3.03 cM. OPN11<sub>980/1070</sub> was clonedand sequenced, and specific PCR primerswere designed to convertOPN11<sub>980/1070</sub> into sequencecharacterized amplified region (SCAR) makerSCN11<sub>980/1070</sub>. SCAR analysis of theF<sub>2</sub> population confirmed thatOPN11<sub>980/1070</sub> and SCN11<sub>980/1070</sub> areat the same locus linked to the SMVresistance gene. The RAPD markerOPN11<sub>980</sub> was used as RFLP probefor southern hybridization to soybeangenomic DNA. Southern analysis showed thatsoybean genome contains low-copy sequenceof OPN11<sub>980</sub>. Using a recombinant inbredmapping population of `Kefeng No.1' (R) ×Nannong1138-2'(S), OPN11_980/1070 was mapped to thesoybean molecular linkage group (MLG) Fbetween the restriction fragment lengthpolymorphism (RFLP) markers B212 (0.7 cM) and K07 (6.7 cM) and 3.03 cM apart from theSMV resistance gene.     

Inheritance of resistance to the Moroccan watermelon mosaic virus in the cucumber line TMG-1 and cosegregation with zucchini yellow mosaic virus resistance

Inbred lines derived from the Chinese cucumber (Cucumis sativus L.) cultivar, ‘Taichung Mou Gua’ (TMG), have been shown to be resistant to several potyviruses including: zucchini yellow mosaic virus (ZYMV), zucchini yellow fleck virus (ZYFV), watermelon mosaic virus (WMV) and the watermelon strain of papaya ringspot virus (PRSV-W). Recently, an additional virus that infects cucurbits, the Moroccan watermelon mosaic virus (MWMV), has been determined to be a distinct member of the potyvirus group. This study demonstrates that TMG-1 possesses resistance to MWMV. Rub or aphid inoculated TMG-1 seedlings remain free of symptoms. Progeny analyses of the F₁, F₂ and backcross generations show that resistance to MWMV is conferred by a single recessive gene (proposed designation, mwm). Sequential inoculation of progeny possessing resistance to ZYMV followed by MWMV (or MWMV followed by ZYMV) and analysis of F₃ families derived from F₂ individuals selected for resistance to ZYMV indicate that both resistances are conferred by the same gene, or two tightly linked genes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Wheat streak mosaic virus resistance in eight wheat germplasm lines

Wheat streak mosaic virus (WSMV) is a destructive pathogen in wheat (Triticum aestivum L.). Host resistance is the most effective way to control this virus. To date, Wsm2 is the only wheat resistance gene that is genetically mapped. The objective of this study was to identify germplasm lines that might carry resistance genes different from Wsm2. Eight newly reported resistant germplasm lines were examined by allelic tests. To validate the allelic test results, five of them were further analysed for the inheritance of WSMV resistance. A Wsm2‐linked marker was also genotyped on populations developed for the inheritance study. Our results suggested that the WSMV resistance in lines CItr9358, PI225288, PI243652, PI245439, PI245526 and PI478095 was controlled by either Wsm2 or a gene very closely linked to Wsm2. The resistance in PI243753 and PI321730, however, is likely controlled by a gene different from, but linked to Wsm2. The resistance in PI321730 might also involve some minor genes. This study provided useful information for breeders to select appropriate resistant lines to improve WSMV resistance in wheat.     

Resistance of Cucurbita moschata to watermelon mosaic virus type 2 and its genetic relation to resistance to zucchini yellow mosaic virus

Summary Cucurbita moschata Menina originating from Portugal is known to carry a single dominant gene, Zym, for zucchini yellow mosaic virus (ZYMV) resistance. Resistance to watermelon mosaic virus type 2 (WMV2) was found in the same cultivar. In resistant plants, WMV2 migration from inoculated leaves or cotyledons seems to be limited and blocked. Resistance to WMV2 is conferred by one dominant gene and is effective against eight strains from different geographic origins. Results of linkage studies, based on co-inoculation of plants with WMV2 and ZYMV, indicate that resistance to these two viruses is conferred by the same gene, probably Zym, or perhaps by two closely linked genes.     

Inheritance and linkage analysis of resistance to zucchini yellow mosaic virus, watermelon mosaic virus, papaya ringspot virus and powdery mildew in melon

A melon (Cucumis melo L.) breeding line derived from PI 414723 is resistant to three potyviruses,watermelon mosaic virus (WMV), zucchini yellow mosaic virus (ZYMV), papaya ringspot virus (PRSV), and to powdery mildew (PM). The inheritance and linkage relationships of these four resistances were studied in a segregating F2 population and derived F3 families from a cross between cultivar Top Mark and the resistant breeding line. Dominant monogenic inheritance of all four resistances was observed. We report that line 414723-4S3, which was initially selected as a source of ZYMV and WMV resistance, is also a source of dominant monogenic resistances to PRSV and PM race 1. We also report on genetic linkage (significant departure from independent segregation, χ² = 58.1, p≪ 0.0001) between resistance to WMV and ZYMV. The map distance between these loci was estimated to be 7.5 cm. The genes for resistance to PM and PRSV segregated independently from each other, and from ZYMV and WMV resistance. This revised version was published online in July 2006 with corrections to the Cover Date.     

侵染南瓜的西瓜花叶病毒和黄瓜花叶病毒CP基因的克隆和序列分析

采用RT-PCR的方法,用马铃薯Y病毒属病毒3′-末端序列的简并引物和黄瓜花叶病毒（Cucumber mosaic virus, CMV）外壳蛋白（CP）基因的特异引物,对采自山东聊城的一表现花叶、黄化、蕨叶及果实畸形的南瓜样品进行了检测,同时扩增到了西瓜花叶病毒（Watermelon mosaic virus, WMV）和CMV 2种病毒的基因组片段,说明该样品受到WMV和CMV 2种病毒的复合侵染。这2个病毒分离物分别被命名为WMV-liaocheng和CMV-liaocheng。序列测定及分析结果表明,它们与其它相应病毒分离物CP基因核苷酸序列的同源性分别为91.2-98.0%和77.0-97.9%,推导的氨基酸序列同源性分别为96.4-98.5%和81.2-99.1%。根据完整CP 基因核苷酸序列构建的系统进化树显示：18个WMV分离物可分为3组,其中WMV-liaocheng 与HLJ、CHN及Habenaria等分离物表现出较近的亲缘关系,形成Ⅲ组;30个CMV分为两个亚组,其中CMV-liaocheng属于亚组I,CMV-liaocheng可能发生过重组。     

Linkage relationship between two genes conferring resistance to peanut stripe virus and soybean mosaic

Summary The possible linkage relationship between hypocotyl color, leaflet shape, resistance to soybean mosaic virus (SMV) and to a soybean isolate of peanut stripe virus (PStV-isolate PN) was examined in two soybean lines AGS 129 and Ankur.Hypocotyl color, leaf shape and reactions to SMV-G1 and PStV were found to be inherited monogenically, with purple hypocotyl color, ovate leaf shape and resistance to both of the viruses being dominant. The reactions to SMV and PStV were conditioned by genes with 9 ± 2.4 percent recombination as coupling phase. They were inherited independently from hypocotyl color and leaf shape.     

甘蓝抗芜菁花叶病毒育种研究进展

摘要：本文简述了甘蓝TuMV抗性种质资源的现状和各种分子生物技术应用发展的前景,从TuMV的发生、甘蓝抗TuMV种质资源的挖掘,甘蓝对TuMV的抗性规律,植-病互作分子机制,甘蓝抗TuMV分子标记的开发与应用和基因工程技术在甘蓝抗TuMV育种中的应用等方面综述了近年来的研究进展,为今后甘蓝抗性育种提供参考和借鉴。     

Resistance of early-maturing European maize germplasm to sugarcane mosaic virus (SCMV) and maize dwarf mosaic virus (MDMV)

Sugarcane mosaic virus (SCMV) and maize dwarf mosaic virus (MDMV) are the most important viruses of maize in Europe. In field and greenhouse experiments, 122 early-maturing European maize inbreds (45 flint and 77 dent lines) were evaluated for their reaction to artificial inoculation by SCMV and MDMV. Three dent inbreds (D21, D32, FAP1360 A) with complete resistance and four dent inbreds (D06, D09, R2306, FAP1396A) with partial resistance against both potyviruses under both greenhouse and field conditions were identified. All other inbreds were highly susceptible to both SCMV and MDMV. Selection for virus resistance in maize breeding could be performed with only one virus at a time because all inbreds resistant to SCMV were also resistant to MDMV. Rank correlations between percentages of infected plants in greenhouse and field trials ranged from 0.51 to 0.72 for both SCMV and MDMV, suggesting that prescreening of breeding materials for virus resistance can be performed in the greenhouse but final evaluation in multilocation trials in the field is recommended.     

Single-gene resistance to zucchini yellow mosaic virus in Cucurbita moschata

Summary The mode of inheritance for resistance to zucchini yellow mosaic virus (ZYMV) in Cucurbita moschata was determined from F₁, F₂, and backcross progenies of the cross between the susceptible cultivar Waltham Butternut from the U.S.A. and a resistant inbred line of the Menina cultivar from Portugal. Resistance to ZYMV in Menina was conferred by a single dominant gene designated Zym.     

Components of resistance to ryegrass mosaic virus in a clone of Lolium perenne and their strain-specificity

Summary The extreme resistance to ryegrass mosaic virus (RMV) of a clone of Lolium perenne L. was due to a combination of two distinct types of resistance: resistance to infection and resistance to multiplication and movement of virus within the plant. Resistance to infection was quantitatively inherited and highly effective against three strains of RMV, while resistance to multiplication and movement was controlled by two complementary recessive genes and was effective against only two of the three RMV strains.     

Characterization of broad‐spectrum resistance to Soybean mosaic virus in soybean [Glycine max (L.) Merr.] cultivar ‘RN‐9’

Soybean mosaic virus (SMV) can cause serious yield losses in soybean. Soybean cultivar ‘RN‐9’ is resistant to 15 of 21 SMV strains. To well‐characterize this invaluable broad‐spectrum SMV‐resistance, populations (F₁, F₂ and F_2:3) derived from resistant (R) × susceptible (S) and R × R crosses were tested for SMV‐SC18 resistance. Genetic analysis revealed that SC18 resistance in ‘RN‐9’ plus two elite SMV‐resistant genotypes (‘Qihuang No.1’ and ‘Kefeng No.1’) are controlled by independently single dominant genes. Linkage analysis showed that the resistance of ‘RN‐9’ to SMV strains SC10, SC14, SC15 and SC18 is controlled by more than one gene(s). Moreover, Rsc10‐r and Rsc18‐r were both positioned between the two simple sequence repeats markers Satt286 and Satt277, while Rsc14‐r was fine‐mapped in 136.8‐kb genomic region containing sixteen genes, flanked by BARCSOYSSR_06_0786 and BARCSOYSSR_06_0790 at genetic distances of 3.79 and 4.14 cM, respectively. Allelic sequence comparison showed that Cytochrome P450‐encoding genes (Glyma.06g176000 and Glyma.06g176100) likely confer the resistance to SC14 in ‘RN‐9’. Our results would facilitate the breeding of broad‐spectrum and durable SMV resistance in soybeans.     

Inheritance of resistance to turnip mosaic virus in Chinese cabbage

Summary Turnip mosaic virus (TuMV) is the most important virus of commercially grown cole crops in many Asian countries, affecting both yield and quality. TuMV-infected Chinese cabbage becomes unmarketable because of the presence of black spots and necrosis often induced by the virus. Resistance breeding is complicated by the existence of five strains of the virus, one of which was discovered in 1985 for the first time in Taiwan. Resistance to strains C1 to C3 is readily available among the Chinese cabbage germplasm at AVRDC, whereas resistance to strains C4 and C5 is rarely found. To elucidate the inheritance of resistance to TuMV, P₁, P₂, F₁, F₂ and BC₁ generations of crosses between the resistant line 0–2 and three susceptible lines, E-7, E-9 and FL-9, were inoculated with strains C4 and C5. Segregation ratios obtained by visual observation and enzyme-linked immunosorbent assay (ELISA) indicate that two recessive genes confer resistance to both TuMV-C4 and TuMV-C5.     

Role of male-sterile cytoplasm in resistance to barley yellow mosaic virus and Fusarium head blight in barley

Two types of male‐sterile cytoplasm, designated msm1 and msm2, in barley were investigated to determine whether these cytoplasms confer resistance to barley yellow mosaic virus (Ba YMV) and Fusarium head blight (FHB). Alloplasmic lines and isogenic lines of two cultivars showed the same reaction to each Ba YMV as that of their euplasmic lines. This demonstrates that the barley male‐sterile cytoplasms msm1 and msm2 have no effect on resistance to BaYMV. No significant difference in reactions to FHB was recognized among fertile alloplasmic lines of ‘Adorra’, but the difference in reactions to FHB between fertile and sterile isogenic lines of ‘Adorra’ was significant. The damage caused by FHB in the male‐sterile lines that produced sterile pollen was significantly greater than the damage in a sterile line that did not produce pollen. These results suggest that pollen or anthers are important factors in infection with or spread of FHB. For production of hybrid seeds, male‐sterile lines with no pollen production, such as those with msm1 male‐sterile cytoplasm, would reduce FHB infestation.     

HC-Pro基因片段介导的高抗TuMV

芜菁花叶病毒(turnip mosaic viruses,TuMV)是侵染重要经济作物的主要病毒之一,寄主范围十分广泛,尤其是对十字花科作物的危害最为严重。为了获得对TuMV持久稳定的高度抗性,本研究以TuMV HC-Pro基因的453 bp保守序列为靶标,构建了植物表达RNAi载体pBBBTu-HC-Pro,并转化了TuMV的天然宿主拟南芥。对转基因拟南芥用北京地区流行的TuMV强致病株BJ-C4进行了抗病接种鉴定。鉴定的13个单拷贝转基因株系中有4个对TuMV的BJ-C4株表现出高度抗性,抗性比对照提高约80%以上,且抗性可以稳定遗传。经半定量和荧光定量PCR方法检测,在高抗转基因植株体内几乎检测不到病毒的累积,抗病效果明显。该载体在利用基因工程抗TuMV育种中具有广阔的应用前景。