InDel分子标记WC656的开发及其在鉴别小麦-簇毛麦5VS纯合易位中的应用

簇毛麦是小麦品种改良重要的遗传资源,其5VS上含有硬度基因Dina/Dinb、抗白粉病基因Pm55和抗条锈病基因Yr5V,创制普通小麦-簇毛麦5VS易位系新种质,对于高效利用5VS上的有益基因进行小麦品种遗传改良具有重要意义。为了便于鉴别普通小麦-簇毛麦5VS纯合易位,本研究以mInDel软件分析普通小麦中国春基因组序列,开发了小麦第5同源群短臂的InDel特异分子标记WC656,对小麦品种以及普通小麦-簇毛麦5VS.5AL和5VS.5DL易位系及其衍生后代进行PCR扩增,根据扩增片段大小来区分5AS、5BS、5DS染色体臂以及5VS易位系。结果表明,在21个普通小麦品种、小麦-簇毛麦5VS回交F₂代中杂合易位单株,均扩增出379 bp（5AS）、471 bp（5BS）、422 bp（5DS）3条带。在T5VS.5AL高代品系、回交F₂代中纯合易位单株扩增出471 bp（5BS）、422 bp（5DS）2条带,379 bp（5AS）条带缺失。T5VS.5DL高代品系、回交F₂代中纯合易位单株扩增出379 bp（5AS）、471 bp（5BS）2条带,422 bp（5DS）条带缺失。WC656鉴别的5VS纯合易位结果与顺序FISH-GISH分析结果一致。T5VS.5AL、T5VS.5DL具有抗白粉病、籽粒硬度指数低等优良特性。因此,利用InDel分子标记WC656可以鉴别普通小麦-簇毛麦T5VS.5AL、T5VS.5DL衍生后代中的纯合易位,PCR扩增实验操作相对简便,可以为分子标记辅助选育携有5VS纯合易位的软质弱筋小麦提供帮助。     

普通小麦川麦60-簇毛麦 3V（3D）代换系的分子细胞学鉴定

一年生二倍体簇毛麦携带多种抗性基因,被认为是小麦抗性改良的重要基因源。本研究前期创制了一个高抗小麦条锈病的普通小麦（中国春,CS）-簇毛麦3V（3D）代换系CD-3,并将该条锈病抗性基因初步定位于簇毛麦3V染色体上。本研究以川麦60作为父本,以CD-3为母本,在回交自交系BC₁F₂群体中筛选到一个高抗条锈病的株系,命名为1901-245,并进一步对该株系进行分子标记、荧光原位杂交（FISH）及田间条锈病抗性鉴定。分子标记和荧光原位杂交（fluorescence in situ hybridization,FISH）结果显示,1901-245携带一对簇毛麦3V染色体,丢失了一对小麦3D染色体;1901-245花粉母细胞减数分裂中染色体配对正常,单价体出现频率较低,且3V染色体未出现单价体。田间条锈病抗性鉴定结果显示,1901-245及其亲本CD-3均表现为高抗小麦条锈病,而亲本川麦60则表现为高感小麦条锈病。以上结果表明,簇毛麦3V染色体上的条锈病抗性基因在1901-245的遗传背景下能正常表达;1901-245为普通小麦-簇毛麦3V（3D）代换系材料,高抗小麦条锈病且遗传稳定,可进一步在小麦遗传改良中加以应用。另外,FISH鉴定及植株形态观察结果显示,1901-245仍含有较多亲本CD-3（CS背景）的遗传背景,因此后期可继续将1901-245与川麦60进行回交,并在后代中加大簇毛麦3V染色体的追踪力度。     

簇毛麦1V染色体臂的EST STS标记的开发与应用

为了给小麦远缘亲本的研究和利用提供参考,根据已定位于普通小麦1DS和1BL上的EST序列设计96对STS引物,对中国春、簇毛麦、中国春-簇毛麦的1V附加系和易位系进行多态性分析,开发出4个簇毛麦1V染色体臂的STS标记。其中,BE499250-STS和BE591682-STS/RsaⅠ为共显性标记,可以扩增出1条1VS片段,同时还可以扩增出1条1DS片断,能将1DS和1AS、1BS区分开来;BE581358-STS/HaeⅢ和BE585781-STS/RsaⅠ是显性标记,能分别扩增出2条和1条1VL片段,但扩增的小麦染色体片段不能区分1DL和1A、1B。这些标记已成功地应用于小麦-簇毛麦整臂互补易位系T1DL.1V#3S和T1DSo1V.3L的筛选。     

簇毛麦基因组特异DNA序列标记的建立和应用

为建立簇毛麦基因组特异DNA序列的分子标记，以普通小麦中国春、绵阳11、川农17和R111以及多年生簇毛麦、硬粒小麦一多年生簇毛麦双二倍体TDB-3为材料，用120条随机引物进行RAPD分析，筛选出一个多年生簇毛麦的特异性RAPD片段。克隆测序表明该片段全长为937bp（记为OPM2 937）。以OPM2 937的DNA序列为基础设计了一对特异引物，并用该特异引物对多年生簇毛麦、二倍体簇毛麦以及中国春-二倍体簇毛麦附加系CSDA1V～CSDA7V与小麦-多年生簇毛麦衍生后代进行了扩增，结果表明，凡具有簇毛麦染色体的材料均可扩增出目标DNA片段，而不具簇毛麦染色体的材料均未能得到扩增。将OPM29 937在NCBI中进行序列比对，发现它是一个新的序列，说明OPM2 937为簇毛麦基因组的一个特异序列，该DNA序列标记可用于快速跟踪检测小麦背景中的簇毛麦染色体。     

普通小麦-簇毛麦1V染色体系的选育与鉴定

为了转移和利用簇毛麦1V染色体上的优质基因,用中国春和硬粒小麦-簇毛麦双倍体杂交,再用中国春回交,综合运用染色体C-分带、荧光原位杂交、高分子量谷蛋白亚基分析和分子标记分析,从BC1F1~BC1F3代中检测到1V染色体系,在BC1F3、BC2F1代中选育出分别涉及簇毛麦1V染色体长臂和短臂的四种染色体结构变异系,包括1VS.W易位系、W.1VL易位系、1VS单端体系和1VL端二体系,为小麦育种创造了新的种质资源。     

簇毛麦6V染色体短臂特异性EST标记的开发及缺失定位

为了准确鉴定簇毛麦6VS易位染色体小片段的长度和断点位置,根据定位于小麦第6部分同源群染色体短臂不同区段的EST序列,利用PRIMER5.0软件设计PCR引物,成功开发出3个对6V染色体短臂特异的分子标记(CINAU89-740、CINAU90-730、CINAU91-390)。利用小麦和簇毛麦第六部分同源群染色体短臂的12个缺失系对簇毛麦6VS的8个特异性分子标记进行了定位。标记CINAU88-381、CINAU17-1086、CINAU15-902分别被定位在FL值为0.79~0.99、0.58~1.00、0.46~0.58的染色体区段,标记CI-NAU91-390、CINAU90-730、CINAU16-1650被定位在FL值为0.35~0.45的染色体区段,标记CINAU18-723、CINAU89-740被定位在FL值为0.00~0.45的染色体区段。利用这8个分子标记可以更准确地鉴定6VS小片段易位的断点和片段长度。     

小麦TaWRKY2基因组分析及在进化和育种材料中的检测

小麦WRKY转录因子在抗旱抗逆等方面有重要作用。为小麦抗旱抗逆分子模块组装育种提供参考,本研究在已有小麦TaWRKY2基因cDNA序列（GenBank登录号：EU665425）的基础上,从小麦品种晋麦79号中克隆获得TaWRKY2基因全长序列,并对TaWRKY2的基因结构、基因组信息进行研究,同时检测其在部分小麦亲缘种、黄淮麦区水旱地代表品种、本课题组选育的高代品系及部分水旱地杂交组合F₁代中的分布。结果表明,克隆所得到的TaWRKY2基因全长DNA序列包含4个外显子和3个内含子,其中,3个内含子长度变化范围为111~172 bp;外显子和内含子的交接处序列符合GT-AG原则;与乌拉尔图小麦（Triticum urartu,AA）和粗山羊草（Aegilops tauschii,DD）中的片段序列同源性分别为93%和99%,与中国春小麦（AABBDD）1AS、1BS和1DS染色体的片段序列同源性分别为93%、88%和99%。除野生一粒小麦外,在小麦进化祖先种、黄淮海水旱地推广的代表品种、课题组选育的高代品系及F₁代共78份材料中,都能够检测出该基因的普遍存在,说明该基因可应用于小麦抗旱抗逆分子设计育种中。     

族毛麦基因组及其在小麦改良中的应用研究进展

簇毛麦是小麦的近缘属，是小麦改良重要的基因资源。簇毛麦有益基因向小麦遗传背景的导入，主要利用小麦－簇毛麦双二倍体为桥梁亲本，通过杂交、回交等方式获得小麦异代换系、异附加系、异易位系；并且利用形态标记、细胞学标记、生化标记及分子标记对小麦遗传背景下的染色体组、染色体、染色体臂及片段进行鉴定。本文对这些方面的研究进展进行了综述，对族毛麦可利用基因进行了展望。     

野生二粒小麦抗条锈病基因 YrH52的RGA分子标记

为开发与野生二粒小麦抗条锈病基因YrH52紧密连锁的分子标记,并为该基因的克隆及应用奠定基础,运用RGA (Resistance gene analog) 分子标记法,以 YrH52定位作图F₂群体形成的F₄抗性和感病基因池（Gene pool）及其亲本（抗病材料H52与感病材料Ldn）进行多态性筛选分析,共获得17个RGA分子标记。使用已有的遗传图并进行MultiPoint分析,构建了由与抗性（H）和感病(L) 两个亲本对应的显性位点组成的两个遗传图,即H遗传图和L遗传图。在H图中, YrH52与10个RGA标记,即 X_uhw3, X_uhw17, X_uhw18, X_uhw23, X_uhw36, X_uhw38, X_uhw46, X_uhw59, X_uhw62 和 X_uhw73 紧密连锁, 其中 X_uhw23 标记为共显性分子标记,连锁距离为1.0 cM。在L图中,发现 X_uhw57, X_uhw68, X_uhw189, X_uhw192及其 X_uhw23与 YrH52 相聚成簇（Cluster）。本研究结果说明RGA分子标记结合集群分离分析法 （Bulked segregant analysis,BSA）是一种快速开发与小麦抗病基因紧密连锁标记的有效方法,对小麦抗条锈病分子育种和抗病基因的克隆具有促进作用。     

67份美国小麦品种矮秆基因的分子标记检测

为了明确矮秆基因在美国小麦品种中的分布特点并发掘较少利用的矮秆基因,本研究利用8个小麦矮秆基因的特异分子标记对67份美国小麦品种中的矮秆基因进行了检测。结果表明,67份美国小麦品种中,超过80%的品种（56个）含有矮秆基因,其中 Rht-B1b和 Rht8基因频率较高,分别占62.7%和43.3%。仅有5个品种含有 Rht-D1b基因,同时发现3个品种含有 Rht5基因,2个品种含有 Rht12基因,未发现含有 Rht4、 Rht9和 Rht13基因的品种。在含有矮秆基因的品种中,35.8%的品种含有2个或2个以上的矮秆基因。其中有1个品种同时含有3个矮秆基因,有20个品种同时含有 Rht-B1b和 Rht8基因,有3个品种同时含有 Rht-D1b和 Rht8基因,有1个品种同时含有 Rht-D1b和 Rht12基因,其余32个品种各含有1个矮秆基因。本研究未发现同时含有 Rht-B1b、 Rht-D1b、 Rht8以及同时含有 Rht-B1b、 Rht-D1b的品种。     

Extreme resistance to infection by potato virus X in genotypes of wild tuber-bearingSolanum species

Summary Clones derived from thirty-one different accessions (nineteen of Argentine origin) belonging to eightSolanum species were screened for resistance to infection by potato virus X strain cp (PVX cp) by mechanical inoculation of plantlets that had been micropropagated in vitro. Estimates of PVX multiplication obtained by enzyme linked immunosorbent assay and slot blot nucleic acid hybridization allowed the identification of resistant clones derived from five accessions belonging toS. commersonii S. oplocense, S. sparsipilum andS. tuberosum andigena. Resistant genotypes supported PVX concentrations 5 to 15 times smaller than did the susceptible control cultivar Spunta. Graft inoculation test confirmed the presence of extreme resistance similar to that conferred by the ‘immunity’ gene X¹ (also called RX_act).     

Effect of a continuous hot water treatment of potato tubers on seed-borne fungal pathogens

Summary The viability of five pathogens was decreased by treatment with hot water when tested in vitro.Polyscytalum pustulans was most sensitive andRhizoctonia solani least sensitive. Potato tubers were exposed to 55°C for 5 min in a commercial continuous hot water treatment plant using naturally contaminated seed tubers and tubers which had been inoculated by dipping in comminuted cultures. The frequency of eyes colonised byP. pustulans, Helminthosporium solani, andR. solani was reduced to virtually zero and the effect persisted on tubers subsequently stored at 4°C and at 15°C for up to 16 wk. Results withColletotrichum coccodes were inconclusive. Treatment suppressedPenicillium spp. which, however, rapidly recolonised the eyes during storage, leading to higher contamination levels in the treated than in the untreated tubers. With tubers inoculated withPhoma foveata, good control was achieved when the incubation period before treatment was 10 d but not when the period was 42 d.     

In vitro antimicrobial activity of different accessions ofSolanum commersonii Dun. from Uruguay

Summary The in vitro antimicrobial activity of extracts from accessions ofSolanum commersonii Dun. collected in the south of Uruguay was investigated against five microorganisms including the pathogenRalstonia solanacearum. A total of 30 extracts corresponding to organic and aqueous extracts were studied. Interestingly, most of the positive results for growth inhibition were againstR. solanacearum. The extracts were also analyzed for the presence of glycoalkaloids and lectins. Six of the ten aqueous extracts showed lectin presence and a wide variation in the type and amounts of glycoalkaloids, was found. Results indicate that there is no clear relationship between the antimicrobial activity against the five microorganisms screened and the presence or amounts of lectins and glycoalkaloids, traditionally regarded as possible antimicrobial metabolites in theSolanum genus, which indicates the presence of as yet unidentified antimicrobial compounds.     

玉米黑粉菌菌瘤内内生细菌对黑粉菌的冬孢子萌发的影响

利用高通量测序分析明确玉米黑粉菌菌瘤内菌粉培养液中内生细菌种类，结合16SrDNA分析，分离并鉴定在培养液中占比最高的菌株。以溶菌酶及Amp处理玉米黑粉菌冬孢子为对照，显微镜下观察菌瘤内内生细菌对玉米黑粉菌冬孢子萌发的影响。结果表明，在4份样品的菌粉培养液中，肠杆菌属细菌的占比在90%以上。当肠杆菌大量增殖时对黑粉菌的冬孢子萌发产生影响，可引起黑粉菌的冬孢子发生质壁分离，释放原生质体，部分原生质体再生成菌体；也可以使冬孢子壁直接被降解，原生质体萎缩。研究发现，玉米黑粉菌菌瘤内的内生细菌以肠杆菌为主，该内生细菌对黑粉菌的冬孢子萌发产生影响。     

Field assessment of the relative importance of different aphid species in the transmission of potato virus Y

Summary Flying aphids were trapped throughout the summer on a vertical net downwind of a plot of PVY-infected potato plants. Of 6769 individuals caught, 165 transmitted PVY to tobacco test seedlings. Of 119 species or species groups caught, 20 were found to be vectors of which nine had not been recorded previously.Brachycaudus helichrysi, Myzus persicae, Phorodon humuli andAphis species accounted for 90% of transmissions andB. helichrysi alone for 52% of transmissions. The prospects of using this information to assess the amount of virus spread in a potato crop and of forecasting the timing and abundance of the main vectors are discussed.

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Zusammenfassung Im Abwind einer Parzelle mit PVY-infizierten Kartoffelpflanzen (7% PVY^O, 4% PVY^N, 89% beide) wurden fliegende Blattl?use in zwei vertikalen Netzen (4,4 m × 1,4 m mit hexagonalen L?chern mit 1,5 mm Durchmesser) gefangen und anschliessend auf Tabaks?mlinge überführt, um zu prüfen, ob sie PVY übertragen. Die Blattl?use wurden vom 8. Juni bis zum 8. August 1984 normalerweise an einem Tag pro Woche von 10 bis 17 Uhr gefangen. Im Laufe von 12 Fangtagen wurden 6769 sachte 52% der übertragungen bei nur 15% der totalen Fangzahl (Tab. 3).B. helichrysi mitMyzus persicae, Phorodon humuli und Species des GenusAphis ergaben sich als 90% der übertragenden Individuen bei nur 32% der gefangenen Proben. Acht Prozent der gefangenenB. helichrysi waren übertr?ger, dagegen nur 4,7% vonM. persicae. B. helichrysi k?nnte als Vektor besonders wichtig sein, weil sie in der Vegetationszeit Blattl?use von 119 Species oder Speciesgruppen gefangen, von denen 165 (2,44%) Tabak mit PVY inokulierten (Tab. 1). 133 Blattl?use übertrugen PVY^O und 32 übertrugen PVY^N (und m?glicherweise auch PVY^O). 20 Species oder Speciesgruppen waren Vektoren, von denen 9 (Cryptomyzus ballotae, Hyperomyzus lactucae, Metopolophium festucae, Myzus ligustri, M. myosotidis, Myzaphis rosarum, Sitobion avenae, S. fragariae undUroleucon spp.) erstmalig beschriebene Vektoren sind.Brachycaudus helichrysi verurfrüh auftritt (Tab. 2), wenn die Pflanzen am anf?lligsten sind und die Virus-Translokation zu den Knollen am schnellsten stattfindet. Die Aussichten zum Gebrauch dieser Information zur Absch?tzung des Aufbaues von Virusbefall im Bestand und die Aussichten für eine Vorhersage der Menge der wichtigsten Vektoren werden diskutiert. Es ist dringend erforderlich, die Biologie vonB. helichrysi zu untersuchen.

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Résumé Deux filets verticaux (4,40 m × 1,40 m à mailles héxagonales de 1,5 mm de diamètre) sont placés face au vent pour la capture des pucerons dans une parcelle de plants de pommes de terre contaminés par PVY (7% PVY^O, 4% PVY^N, 89% par les deux) puis les pucerons sont transferrés sur des plantules de tabac en milieu clos pour tester la transmission de PVY. Les captures de pucerons ont eu lieu du 8 juin au 8 ao?t 1984, en général à raison d'un journée par semaine, de 10 heures à 17 heures. Pendant les 12 premiers jours, 6769 pucerons de 119 espèces ou groupes d'espèces ont été capturés, les plantules de tabac ayant été inoculées par PVY avec 165 pucerons (2,44%) (tabl. 1). 133 pucerons étaient vecteurs de PVY^O et 32 de PVY^N (et probablement PVY^O). 20 espèces ou groupes d'espèces étaient des vecteurs dont 9 récemment enregistrés (Cryptomyzus ballotae, Hyperomyzus lactucae, Metopolophium festucae, Myzus ligustri, M. myosotidis, Myzaphis rosarum, Sitobion avenae, S. fragariae etUroleucon spp.).Brachycaudus helichrysi représentait 52% des transmissions mais seulement 15% du total capturé (tabl. 3).B. helichrysi, Myzus persicae, Phorodon humuli et les espèces du gèneAphis représentaient 90% des transmissions et 32% des captures seulement. 8,0% deB. helichrysi capturé a transmis le virus contre, 4,7% pourM. persicae. B. helichrysi peut être particulièrement important comme vecteur dans la mesure ou il appara?t t?t en saison (tabl. 2) lorsque les plantes sont très sensibles et la translocation di virus aux tubercules la plus rapide. Les perspectives à partir de cette connaissance sont discutées en vue d'analyser l'évolution du virus dans une culture et les prévisions de l'abondance des plus importants vecteurs. Une plus grande connaissance de la biologie deB. helichrysi devient urgente.

     

Mineral composition of some traditional Mexican teas

Teas of plant origin traditionally consumed by the Mountain Pima of Chihuahua, Mexico, were analyzed for mineral nutritional content. Fe, Cu, Zn, Ca, and Mg composition was determined for native teas made from shoots ofTagetes lucida, T. filifolia, Elytraria imbricata, andHolodiscus dumosus, and from root xylem ofCeanothus depressus andPhaseolus ritensis. Native uses of these teas are also described.     

Effect of haulm treatment and harvest time on incidence of tuber rots of potato (Solanum tuberosum L.) after standard wounding and on frequency of stem lesions caused byPhoma foveata Foister

Summary Various haulm treatments and harvest times differentially influenced the incidence of storage rots in potato tubers (Solanum tuberosum L.) in field experiments in Sweden performed over 4 years. The incidence of gangrene caused byPhoma foveata Foister (Phoma exigua var.foveata (Foister) Boerema) increased in cv. Bellona after spraying haulms with diquat as did the amount of infection caused byFusarium avenaceum (Corda ex. Fr.) Sacc. in all three cultivars (Bintje, Bellona, and Magnum Bonum). The lowestP. foveata infection levels occurred after pulling haulms. Late harvesting increased the frequency ofP. foveata in cvs Bintje and Bellona both at 2 and 4 wks interval between haulm treatment and harvest. Tuber and visible stem infection caused byP. foveata were correlated for cvs Bintje and Bellona, but not for cv. Magnum Bonum. The possible relation between tuber and stem infection is discussed.     

先玉335、郑单958和京科968抗虫能力的比较研究

以先玉335、郑单958、京科968为材料,以玉米螟(Ostrinia furnacalis)、黏虫(Mythimna separata)和禾谷缢管蚜(Rhopalosiphum padi)为供试昆虫,探究玉米品系大喇叭口期对玉米螟和黏虫及抽雄期对禾谷缢管蚜抗性差异,测定其主要抗虫化合物丁布类物质的含量。结果发现,京科968对玉米螟及禾谷缢管蚜抗性均高于另外两个品系;对黏虫的抗性,郑单958和京科968高于先玉335。大喇叭口期京科968中丁布类物质含量最高,抽雄期郑单958和京科968中主要丁布成分含量均高于先玉335,上述组织中丁布合成相关基因在郑单958及京科968中的表达量也均高于先玉335。本研究揭示杂交玉米抗虫性与抗性相关化合物含量及其相关基因表达间存在正向相关关系,为抗虫玉米分子育种提供理论参考。     

Evaluation of fungicides to control Petri disease pathogens in the grapevine propagation process

The efficacy of fourteen selected fungicides against Phaeomoniella chlamydospora and Phaeoacremonium aleophilum was evaluated in vitro by mycelial growth and conidial germination assays. Azoxystrobin, carbendazim and tebuconazole were the most effective fungicides against P. chlamydospora, while carbendazim and didecyldimethylammonium chloride were the most effective against P. aleophilum. An assay was also conducted with cubiet, hydroxyquinoline sulphate and didecyldimethylammonium chloride to determine their potential to prevent infections caused by these pathogens during the hydration stage in grapevine propagation. The results of this experiment demonstrated that P. chlamydospora and P. aleophilum were able to infect healthy cuttings during the hydration stage and showed that didecyldimethylammonium chloride was consistently the best treatment as a disinfectant soak for grapevine cuttings. Additionally, semi-commercial trials were conducted in two grapevine nurseries to evaluate the effectiveness of several strategies with carbendazim, flusilazole and hydroxyquinoline sulphate at different nursery stages. Soaking planting material in carbendazim during the hydration stage was the most effective treatment since neither P. chlamydospora nor P. aleophilum were isolated from planting material in either nursery.     

The effect of the moisture level of potting compost on the susceptibility of glasshouse-grown tubers to late blight

Summary The effect of the moisture level of potting compost at harvest on the susceptibility of tubers of three cultivars to late blight was investigated in the glasshouse in three successive years. Three moisture regimes (dry, moist, wet) were imposed at flowering time, the tubers harvested 3 weeks later and immediately dipped in a zoospore suspension ofPhytophthora infestans. Tubers from dry compost were significantly more susceptible than those from wet or moist compost, which showed similar levels of susceptibility.