三个小麦野生近缘种抗条锈性传递的初步研究

以当前小麦条锈病菌的优势小种条中２９ 号、３０ 号和３１ 号测定了小麦近缘植物长穗偃麦草、簇毛麦和华山新麦草及其各自与小麦的杂交后代的抗条锈性。试验结果表明,３ 个小麦近缘植物均含有宝贵的抗条锈基因,此类基因具有较强的传递性能,可在小麦遗传背景下高度表达,表现出良好的抗条锈性能,具有广阔的应用前景     

三个小麦野生近缘种抗条锈性传递的初步研究

 以当前小麦条锈病菌的优势小种条中29号、30号和31号测定了小麦近缘植物长穗偃麦草、簇毛麦和华山新麦草及其各自与小麦的杂交后代的抗条锈性。试验结果表明,3个小麦近缘植物均含有宝贵的抗条锈基因,此类基因具有较强的传递性能,可在小麦遗传背景下高度表达,表现出良好的抗条锈性能,具有广阔的应用前景。     

番茄叶霉病高抗基因Cf-9、Cf-11和Cf-19的分子标记

 本研究以9个含不同叶霉病抗病基因的番茄品种为试材,通过接种鉴定表明,Cf-5、Cf-9、Cf-11和Cf-19基因对我国目前的2个叶霉菌优势生理小种均具有较强的抗性。根据Cf-9基因设计引物,扩增Cf-9基因的片段,含Cf-9、Cf-11和Cf-19基因的3种番茄均获得了2.7kb的扩增片段。但用限制性内切酶TaqⅠ对PCR产物酶切可以将3种材料明显区分开来,Cf-9的2个差异酶切片段为1170和460bp;Cf-11的2个差异酶切片段为1100和410bp;Cf-19的2个差异酶切片段为1210和300bp,从而建立了3个基因的分子标记。在F₂分离群体中验证表明,3个基因的分子标记鉴定结果与抗性接种鉴定结果是一致的,用这些标记可以进行分子标记辅助选择。     

源于柔软滨麦草抗小麦条锈病基因YrElm的遗传分析与SSR标记

 M852-1是由柔软滨麦草和普通小麦7182经杂交和回交培育的易位系。苗期抗病性鉴定结果表明,M852-1对CYR29、CYR31、CYR32、CYR33、Su11-4、Su11-7和V26等7个中国小麦条锈菌主要生理小种或新的致病类型均表现免疫至高抗,是一个较好的抗条锈资源材料。用条锈菌流行小种CYR33对M852-1与铭贤169杂交F₁、F₂、F₃和BC₁代进行抗性鉴定与遗传分析,发现M852-1对CYR33的抗条锈性由1对隐性基因控制,暂定名为YrElm。以F₂代分离群体构建作图群体,利用集群分离分析法,筛选到与YrElm连锁的5个SSR标记:Xcfd35、Xgwm161、Xwmc630、Xgwm533和Xcfd34,并将YrElm定位于小麦染色体3DS上。YrElm两侧最近2个SSR标记Xcfd35与Xgwm161的遗传距离分别为6.5 cM和4.2 cM。抗锈性鉴定、系谱分析以及分子标记检测结果表明,该抗病基因来源于柔软滨麦草。综合基因来源、分子检测及染色体位点等方面的分析,认为YrElm可能是一个新的抗条锈病基因。用该基因两侧最近两个标记Xcfd35和Xgwm161 检测68个甘肃和黄淮麦区小麦品种(系),10个(14.7%)品种能扩增出与M852-1相同的条带。进一步进行抗病性及系谱分析表明,这10个品种均不含YrElm。本研究结果为利用YrElm进行分子标记辅助育种和进一步的精细定位奠定了基础。     

结球甘蓝抗TuMV基因的RAPD和SCAR标记研究

 One hundred and forty-four F₂ individuals from a cross combination between 1047 (susceptible) and A21 (resistant) were analyzed for the presence of random amplified polymorphic DNA (RAPD) markers linked to Turnip mosaic virus (TuMV) resistant gene in cabbage by using bulked segregant analysis (BSA).Two polymorphic markers were screened out of 200 random primers.These two RAPD fragments were linked to the resistant gene at 7.7 cM and 8.38 cM respectively and converted to SCAR markers successfully.     

116个小麦品种（系）抗叶锈基因Lr9-Lr26、Lr19-Lr20的复合PCR检测

[目的]建立简单、快速、有效的小麦抗叶锈基因复合PCR体系,从而提高分子标记辅助选择效率。[方法]以28个‘Thatcher’为背景的近等基因系和16个已知基因载体品系作为试材,测试了小麦抗叶锈病基因Lr9、Lr26、Lr19和Lr20的STS标记特异性,通过优化PCR反应体系和循环条件,构建了抗叶锈基因Lr9-Lr26和Lr19-Lr20的复合PCR检测体系。对116个小麦品种(系)所含有的抗叶锈病基因进行了分子检测。[结果]供试品种均不含有Lr9和Lr20,47个品种含有Lr26(基因频率为40.5%),‘中梁22’含有Lr19。经反复验证,Lr9-Lr26和Lr19-Lr20复合PCR技术检测结果可靠,且与上述单个分子标记检测结果一致。[结论]建立的Lr9-Lr26和Lr19-Lr20的复合PCR检测体系可以准确、稳定、高效地检测小麦抗叶锈基因Lr9、Lr26、Lr19和Lr20。     

A SCAR marker specific for rapid detection of the avirulence gene Avr1c in Phytophthora sojae

The F₂ population derived from a cross between isolates pR_x (Avr1c-Avr1c) and ps₁ (avr1c-avr1c) of Phytophthora sojae, fungal agent of soybean stem and root rot, was used to determine the genetic basis of avirulence towards Rps1c gene in soybean. The results indicated that this avirulence is dominant and controlled by a single locus, as expected for a simple gene-for-gene model. Segregation of Avr1c in the F₂ progeny of this cross fits a 3:1 ratio. Four of 80 AFLP primers effectively distinguished the avirulent pR_x from the virulent ps₁. Among the 5 specific markers, band C was amplified from the avirulent pR_x by primer set EGC/MAT, then recovered and cloned. This AFLP marker was successfully transfered to a SCAR marker through sequencing, primer design and specific amplication of the DNA of the avirulent pR_x. Results of validity and specificity experiments with 50 individuals of the F₂ progeny and 50 field isolates demonstrated that this SCAR marker (a 616-bp fragment) can be successfully and specifically amplified from the P. sojae isolates that have Avr1c gene.     

稻曲病菌的SCAR标记及其PCR检测

为建立稻曲病菌Ustilaginoidea virens快速、灵敏的PCR早期检测技术,以S380为引物,对稻曲病菌和其它参试病原菌的DNA进行RAPD-PCR扩增。稻曲病菌RAPD特异片段经回收、克隆和测序后,根据序列用Primer 5.0软件设计了特异性SCAR引物US-SF（5’-TCGCCTCAGACCTCAATC-3’）/US-SR（5’-GAGCCTCAAATGCCTTCC-3’）和巢式PCR引物US-NF（5’-AGCGTCTCCTGCAACC-AC-3’）/US-NR（5’-GAGCCTCAAATGCCTTCC-3’）。利用引物US-SF/US-SR通过常规PCR对供试稻曲病菌均可扩增出1条大小约257 bp的清晰条带,对其它植物病原菌DNA的PCR产物均无扩增条带,最低可检测到1 pg/μL 的病菌基因组DNA;而利用引物US-SF/US-SR和US-NF/US-NR通过巢式PCR可从10 fg/μL的病菌基因组DNA中扩增出1条大小约210 bp的特异性条带。试验表明,巢式PCR检测灵敏度比常规PCR提高了100倍,且巢式PCR可从接菌的水稻幼颖和自然感染的谷粒中检测出稻曲病菌。     

Genetic diversity of the root‐knot nematode Meloidogyne ethiopica and development of a species‐specific SCAR marker for its diagnosis

Meloidogyne ethiopica is an important nematode pathogen causing serious economic damage to grapevine in Chile. In Brazil, M. ethiopica has been detected with low frequency in kiwifruit and other crops. The objectives of this study were to evaluate the intraspecific genetic variability of M. ethiopica isolates from Brazil and Chile using AFLP and RAPD markers and to develop a species‐specific SCAR‐PCR assay for its diagnosis. Fourteen isolates were obtained from different geographic regions or host plants. Three isolates of an undescribed Meloidogyne species and one isolate of M. ethiopica from Kenya were included in the analysis. The results showed a low level of diversity among the M. ethiopica isolates, regardless of their geographical distribution or host plant origin. The three isolates of Meloidogyne sp. showed a high homogeneity and clustered separately from M. ethiopica (100% bootstrap). RAPD screenings of M. ethiopica allowed the identification of a differential DNA fragment that was converted into a SCAR marker. Using genomic DNA from pooled nematodes as a template, PCR amplification with primers designed from this species‐specific SCAR produced a fragment of 350 bp in all 14 isolates of M. ethiopica tested, in contrast with other species tested. This primer pair also allowed successful amplification of DNA from single nematodes, either juveniles or females and when used in multiplex PCR reactions containing mixtures of other root‐knot nematode species, thus showing the sensitivity of the assay. Therefore, the method developed here has potential for application in routine diagnostic procedures.     

四川省17个主栽小麦品种中Lr26和Lr19检测

正小麦叶锈病是一种重要的小麦病害,培育和种植抗病品种是最为经济有效的防治方法。我国在小麦抗病育种工作中引入了1BL/1RS易位系,已知小麦抗病基因Lr26、Pm8、Yr9和Sr31均位于1BL/1RS易位系上~([1])。然而目前Lr26已丧失抗性~([2]),所以育种过程中需要谨慎考虑Lr26的使用。Lr19来源于长穗偃麦草~([3]),在我国目前为有效的     

小麦条锈菌水源11类群的RAPD分析及SCAR标记的建立

鉴于水源11类群近年来一直处于优势地位,为简化其检测手段,本研究利用RAPD技术对该类群的8个主要致病类型进行了多态性分析,以寻找其中主要流行类型的特异性分子标记。结果如下:共筛选出10个碱基随机引物190条,其中94条可得到稳定清晰的扩增图谱,用该94条引物进行RAPD分析,发现各致病类型间遗传变异丰富;以引物S1410扩增得到了水源11-4的特异性DNA条带;以引物S1412和S1304扩增得到了水源11-14的特异性DNA条带;对引物S1304扩增得到的特异性DNA条带回收、克隆和测序,设计了1对19bp/18bp的引物,并成功地将其转化为对水源11-14特异的SCAR标记。以上结果表明,通过规模筛选来寻找小麦条锈菌生理小种的特异性DNA片段,并将其转化为稳定的SCAR标记,有可能建立起中国小麦条锈菌流行生理小种的快速分子鉴定体系。     

小麦条锈菌新菌系V26的SCAR检测标记

 建立小麦条锈菌生理小种的快速分子检测技术体系对小麦条锈菌的监测和防治策略的制定具有重要价值。条锈菌V26是近年来出现的,对我国目前小麦抗病育种中普遍应用的抗条锈病基因Yr26具有毒性的新菌系。该菌系的出现,对我国当前小麦生产、抗病育种都造成了严重威胁。本研究选用189条随机引物对CYR29、CYR31、CYR32、CYR33、T4、Su11-4和V26等7个条锈菌生理小种(菌系)进行了扩增,筛选V26的特异性RAPD片段,并对其进行克隆和测序。根据测序结果,设计并合成SCAR特异性引物, 将V26的RAPD标记转化为稳定的SCAR标记。使得对该菌系的快速检测成为可能,同时也将会为条锈菌新小种的监测提供更为准确的科学依据。     

Histological studies of the expression of the Lr9, Lr20 and Lr28 alleles for resistance to leaf rust in wheat

The development of the leaf rust fungus ( Puccinia recondita f.sp. tritici ) in a susceptible cultivar and three other cultivars possessing the Lr9, Lr20 and Lr28 alleles for resistance was studied by light and fluorescence microscopy. Formation of the substomatal vesicle, intercellular hypha and the first haustorial mother cell was unaffected by resistance. Lr9 and Lr28 expression was rapid, first seen as early initiation of hyphal branching at 16 h after inoculation, then reduced haustorial diameters at 19 h. Limited host cell necrosis was seen immediately afterwards. Elongation of intercellular hyphae was reduced between 20 and 24 h, and virtually ceased by about 30 h. Numbers of infection sites with a second haustorial mother cell were briefly higher at 24 h. Reduced hyphal branching and haustorial mother cell numbers were seen at 20–24 h and 36 h respectively. Lr20 expression was not seen until 36 h when reduced hyphal branching was observed, accompanied by extensive host cell necrosis. Reduced haustorial mother cell numbers were detected at 48 h. Findings suggested a secondary role for host cell necrosis in the expression of the Lr9 and Lr28 alleles. Host necrosis may play a determinant role in Lr20- based expression.     

白僵菌绿僵菌分生孢子对高温的耐受力

试验研究了２株白僵菌和３株绿僵菌的分生孢子在高温短时处理及高温培养条件下的萌发和成活。３５℃短时处理对孢子的萌发和生长基本无影响。４０～５０℃是孢子的敏感区。从高纬度地区吉林采集的菌株Ｂ９对高温最敏感，在４０℃处理１０分钟，发芽率下降８０％。采自非洲的菌株Ｍ１８９对高温的抵抗力最强，在４０℃处理４小时和５０℃处理１小时，孢子的发芽率保持在４０％以上，６０℃处理２０分钟仍有大约１％的发芽率。高温培养结果表明，３５℃时只有Ｍ１８９的成菌落数为６７．７％，其它４个菌株均在１％以下。     

Interactions between Fusarium oxysporum f. sp. racheiphilum and Meloidogyne spp. in Vigna unguculata. 2. Specificity of different taxa

A California isolate of Meloidogyne javanica increased Fusarium wilt symptoms in cowpea cultivars California Blackeye No. 3 (CB3) (resistant to wilt) and Grant (tolerant) inoculated with each of the three races of Fusarium oxysporum f. sp. tracheiphilum. The same isolate of M. javancia did not similarly increase wilt in wilt-resistant cultivar CB7977 inoculated with two isolates of race 3 of F. o. tracheiphilum. Six of seven isolates of M. javanica caused similar increases in vascular discoloration in cultivar CB3. but one isolate of M. javanica and seven of M. incognita did not. Vascular discoloration rating was positively correlated with galling severity. However, increasing the initial inoculum density, and thus galling index, of one isolate of M. incognita did not increase vascular discoloration. The vascular discoloration ratings for the wilt-susceptible CB5 controls in each experiment were higher than those for the wilt-resistant cultivars infected with M. javanica. It is hypothesized that M. javanica but not M. incognita reduces, but does not eliminate, resistance to all races of F. o. tracheiphilum in cultivars CB3 and Grant.