甘蔗-滇蔗茅杂交F_1花粉母细胞减数分裂过程GISH分析

研究甘蔗属与滇蔗茅属间远缘杂交F1染色体遗传行为具有重要科学意义,可为发掘利用滇蔗茅野生优异基因资源提供细胞学依据。本研究采用常规压片技术对可育父本及不育杂交F1花粉母细胞减数分裂过程进行比较观察,结果显示可育父本云南95-19减数分裂正常,而不育杂交F1分裂异常;进一步对F1花粉母细胞减数分裂过程进行基因组荧光原位杂交(GISH,genome in situ hybridization)分析,结果表明:滇蔗茅与甘蔗属热带种的亲缘关系较远,双亲染色体在F1细胞中不能进行同源配对,终变期,15条滇蔗茅染色体以单价体形式存在,花粉母细胞减数分裂细胞中存在滞后染色体、染色体丢失和不均衡分离现象。甘蔗-滇蔗茅属间远缘杂交F1花粉母细胞减数分裂异常因而杂交F1花粉完全败育。     

甘蔗野生种滇蔗茅种质创新利用研究 Ⅰ. 甘蔗与滇蔗茅远缘杂交F1群体构建与SSR分子标记鉴定

利用光周期调控技术诱导甘蔗热带种路打士开花,并与滇蔗茅云南95-19进行远缘杂交,培育实生苗76株,大田移栽成活62株,成活率81.6%;选用4对引物对获得的62份杂交后代进行SSR分子标记鉴定,结果表明,62份杂交后代全部为真杂种,所选SSR引物对滇蔗茅后代群体具有较高的鉴定效率。该杂交组合后代杂种真实率为100%,是目前数量最大的滇蔗茅F1群体。     

甘蔗优良品种材料抗高粱花叶病毒鉴定与评价

采用生长期人工切茎接种和RT-PCR检测相结合方法,于2011—2012年2次对国家甘蔗体系近年选育的49份优良新品种(系)和19份云蔗系列优良中间材料进行由高粱花叶病毒(SrMV-HH,GenBank登录号DQ530434)引起的甘蔗花叶病的抗性鉴定与评价。结果表明,49份优良新品种(系)中,1级高抗到3级中抗的有29份,占59.18%。其中粤甘40号、粤甘42号、粤糖55号、云蔗03-194、云蔗99-596、云瑞06-189、桂糖30号、德蔗03-83、闽糖01-77等9份材料表现1级高抗,占18.37%;福农0335、柳城05-129、粤糖96-86、云蔗01-1413、云蔗03-258、云蔗06-80、桂糖02-351等7份材料表现2级抗病,占14.29%。19份云蔗系列优良中间材料中,1级高抗到3级中抗的有13份,占68.42%。其中云蔗04-622、云蔗05-197、云蔗06-267、云蔗05-194、云蔗06-160、云蔗07-2384、云瑞05-704等7份材料表现1级高抗,占36.84%;云蔗06-362表现2级抗病,占5.26%。研究结果为深入开展甘蔗抗花叶病育种,选育和推广优良抗病品种,有效防控甘蔗花叶病提供了科学依据和优良抗源材料。     

甘蔗野生种滇蔗茅利用研究 Ⅲ.滇蔗茅杂种F1群体的表型变异与遗传多样性分析

选用8个数量性状和18个质量性状对滇蔗茅杂种F1群体的表型变异、多样性及聚类关系进行分析。结果表明：数量性状的遗传变异主要来自于有效茎、单茎重及糖分;质量性状的叶鞘背毛群与内叶耳形状多样性丰富,根点排列、芽位、茎形、节间性状、生长裂缝等5个性状均表现一致;聚类分析表明62份后代材料可分为4大类群和10个亚类群。遗传多样性和聚类分析为杂交后代的筛选及其杂交或回交利用提供了科学依据。     

甘蔗近缘种滇蔗茅(Erianthus rockii)表型性状遗传多样性研究

选用6个数量性状和23个质量性状对滇蔗茅野生资源的表型变异、多样性及聚类关系进行分析。结果表明,数量性状变异系数在12.72%~22.38%之间,最大的是田间锤度,为22.38%,其次是茎径,为22.27%,最小的是叶长,为12.72%;质量性状的曝光后节间颜色多样性丰富,茎形、芽形、生长裂缝、节间形状、花序形状等5个性状均表现一致;聚类分析表明51份滇蔗茅无性系材料可分为4大类群和5个亚类群。遗传多样性和聚类分析为资源杂交利用和优异基因挖掘提供参考。     

甘蔗与斑茅BC1分子鉴定、抗黑穗病和花叶病初步评价

为了解甘蔗(Saccharum)与斑茅(Erianthus arundinaceus)杂交后代作为抗病亲本的利用价值,通过特异引物PCR鉴定出78份甘蔗与斑茅杂交BC_1真实杂种;通过人工接种花叶病毒和黑穗病菌,初步评价了甘蔗和斑茅杂交BC_1的抗病表现。结果表明,甘蔗和斑茅杂交BC_1的抗花叶病具有普遍性,而黑穗病抗性则出现分离。初步筛选出BC_1无性系YCE01-48、YCE01-71、YCE01-105、YCE01-125、YCE02-184和YCE01-118可同时抗花叶病和黑穗病,有望成为甘蔗杂交利用的高抗病源亲本。     

不同无性系蔗茅抽穗开花特性的观察和花粉贮藏条件的研究

来源于较低海拔地区的3个蔗茅无性系的抽穗期较短,来源于较高海拔(大于1500m)地区的16个蔗茅无性系的抽穗期较长,且抽穗期与原产地海拔呈显著负相关(F=-0.827**).蔗茅的这一特性在集中种植后仍能相对稳定地表现出来.蔗茅花粉量较多,花粉发育良好.在自然条件下,蔗茅新鲜花粉仅能存活1 h左右;而经低温(10℃)硅胶干燥2 h后贮藏于-80℃条件下,其生活力保持可超过360d.蔗茅贮藏花粉生活力的下降比率与花粉含水量呈显著正相关(r=0.933**);含水量降至10%以下的花粉在低温中贮藏时才能保持较高生活力.     

甘蔗与斑茅割手密复合体杂交后代的分子标记鉴定

为有效利用甘蔗野生种质拓宽甘蔗遗传基础,本研究利用甘蔗与斑茅割手密复合体进行杂交,同时应用序列相关扩增多态性(SRAP)和微卫星(SSR)分子标记技术鉴定其后代。两种分子标记鉴定结果相互印证表明:3个杂交组合的后代中,经表型鉴定含斑茅血缘的34个后代为真杂种。该真杂种后代为进一步综合利用斑茅、割手密的优异基因改良甘蔗品种提供了优良的创新种质。     

蔗茅野生种ErDREB1A基因的克隆与表达分析

为了充分利用蔗茅野生资源,培育耐寒转基因甘蔗提供可靠的候选基因,本研究以电子克隆结合RT-PCR的方法,在蔗茅(Erianthus fulvus)野生种(昆明蔗茅99-1)中克隆到1个干旱应答元件结合蛋白(dehy-dration responsive element binding protein,DREB)基因.生...     

蔗茅EfMYB1基因的克隆与表达分析

为了充分利用蔗茅(Erianthus fulvus)野生资源,挖掘其优良的抗性基因,丰富转基因甘蔗育种候选基因库,该研究结合蔗茅转录组数据,以蔗茅99 1无性系为试验材料,利用RT PCR技术克隆蔗茅MYB基因,并对其进行生物信息学分析及胁迫表达分析,以解析蔗茅的耐寒机理,为转基因甘蔗育种奠定理论基础。结果表明：(1)成功克隆得到一个蔗茅MYB基因,命名为EfMYB1基因(登录号ON586646)。(2)生物信息学分析表明,EfMYB1基因全长1 000 bp,ORF为759 bp,编码251个氨基酸;编码蛋白具有一个保守的SANT结构域,无跨膜结构和信号肽,有多个磷酸化位点;二级结构与三级结构主要以α螺旋和无规则卷曲为主;与南荻相似性最高,遗传距离最近。(3)qRT PCR分析结果发现,EfMYB1基因在蔗茅根和叶组织中的相对表达量随低温胁迫时间的持续而逐渐显著上调,并于胁迫72 h时达到最大值,而在茎中的表达则几乎没有变化;茉莉酸甲酯胁迫下,EfMYB1基因的相对表达量呈先升高后降低的趋势,且在处理6 h时达到最高值;脱落酸胁迫下EfMYB1基因的表达水平较0 h时极显著降低。研究认为,EfMYB1基因属于低温胁迫响应基因,可能参与蔗茅低温胁迫下的应答反应。     

Genetic basis of resistance to sugarcane mosaic virus in European maize germplasm

 Sugarcane mosaic virus (SCMV) causes considerable damage to maize (Zea mays L.) in Europe. The objective of the present study was to determine the genetic basis of resistance to SCMV in European maize germplasm and to compare it with that of U.S. inbred Pa405. Three resistant European inbreds D21, D32, and FAP1360A were crossed with four susceptible inbreds F7, KW1292, D408, and D145 to produce four F₂ populations and three backcrosses to the susceptible parent. Screening for SCMV resistance in parental inbreds and segregating generations was done in two field trials as well as under greenhouse conditions. RFLP markers umc85, bnl6.29, umc10, umc44, and SSR marker phi075 were used in F₂ populations or F₃ lines to locate the resistance gene(s) in the maize genome. Segregation in the F₂ and backcross generations fitted to different gene models depending on the environmental conditions and the genotype of the susceptible parent. In the field tests, resistance in the three resistant European inbreds seems to be controlled by two to three genes. Under greenhouse conditions, susceptibility to SCMV in D32 appears to be governed by one dominant and one recessive gene. Allelism tests indicated the presence of a common dominant gene (denoted as Scm1) in all three resistant European inbreds and Pa405. Marker analyses mapped two dominant genes: Scm1 on chromosome 6S and Scm2 on chromosome 3. Received: 17 November 1997 / Accepted: 25 November 1997     

Identification of somaclonal variants of sugarcane (Saccharum spp.) resistant to sugarcane mosaic virus via RAPD markers

Somaclonal variants resistant to sugarcane mosaic virus (SCMV) were obtained from susceptible sugarcane cv PR62258 through somatic embryogenesis by increasing the number of subcultures of the embryogenic callus tissue in MS medium with 3 mg/L 2,4-dichlorophenoxyacetic acid. Transfers were made at 30-day intervals for 1, 2 or 3 subcultures. Two somaclones, namely AT626 and BT627, were selected by their resistance to SCMV. These subclones have maintained the resistance trait over seven years of testing in the field. In this report we identified the somaclonal SCMV resistant variants from the maternal line and the nonresistant somaclones, using the RAPD technique.     

豇豆优异种质综合评价

摘要 以60份具有早熟、矮秆、多荚、大粒、抗逆、抗病虫、高蛋白性状的优异种质为材料,在河北、河南与安徽进行多点农艺性状综合评价,评选出48份优异种质,其中34个品种达到一级优异种质标准.这些优异种质已结合鉴定繁种入国家中期库,以备科研与生产利用.     

甘蔗花叶病毒CP基因的高效表达和抗血清制备

将甘蔗花叶病毒的CP基因克隆到表达载体pET22b( ),并在大肠杆菌BL21(DE3)中得到高效表达,表达蛋白约占总蛋白的15％。用此蛋白制备了效价高专化性强的抗体。此方法可以解决制备马铃薯Y病毒属病毒的血清时遇到的病毒提纯产量低和寄主蛋白影响等问题。     

黄瓜资源对黄瓜绿斑驳病毒广东分离物的抗性水平鉴定

【背景】黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)是严重威胁葫芦科作物生产的毁灭性病原之一,该病毒已入侵我国十多个省份,危害西瓜、黄瓜等作物并造成严重的经济损失。早在2009年广东即发现CGMMV为害西瓜和黄瓜,但黄瓜等葫芦科作物对其抗性情况尚不清楚。【方法】采用人工机械摩擦接种方法,测定了14份黄瓜种质资源对CGMMV广东分离物的抗性水平。【结果】从广东葫芦病样中分离获得CGMMV,该病毒分离物MP基因序列与国内报道的各分离物同源率均在99%以上;14份黄瓜种质资源对该病毒分离物均表现为感病。【结论与意义】广东主要黄瓜资源对CGMMV均表现为感病,这为我省防控该病毒病提供了科学依据,也为黄瓜抗病育种提供了指导。     

玉米种质对黄曲霉菌抗性的鉴定

以145个玉米自交系作为试验材料,对其籽粒人工接种黄曲霉菌(Aspergillus flavus),根据籽粒的发病情况进行抗性鉴定。结果表明:(1)各玉米自交系对黄曲霉菌的抗性有极显著的差异,在145个自交系中,高抗的8份,中抗的72份,中感的57份,高感的8份;(2)对黄曲霉菌有较高抗性的自交系大多数来自于我国的南方地区;(3)通过辐射处理,对于提高抗性可能有较好的效果;(4)爆裂玉米具有较好的抗性;(5)不同致病菌株接种,抗性反应有差异。     

病毒侵染对转SCMV-CP基因甘蔗光合速率和活性氧代谢的影响

对转SCMV-CP基因甘蔗叶绿素含量、光合参数、活性氧代谢有关的酶活性进行了测定分析,结果表明转基因甘蔗由于外源CP基因的导入,病毒在植株体内无法正常复制繁殖,从而保护了植株细胞免受因病毒侵染造成的伤害。未转基因甘蔗接种后病毒迅速繁殖蔓延,受害叶片对活性氧清除能力的下降,促使有毒物质活性氧的积累,启动膜脂过氧化,造成膜的损伤。     

The complete sequence of a sugarcane mosaic virus isolate causing maize dwarf mosaic disease in China

The complete sequence of a potyvirus from maize in Zhejiang Province was determined. The RNA was 9596 nucleotides long, excluding the 3′-poly (A) tail, and there was a single long open reading frame (ORF) of 9192 nts encoding a 346.1 ku polyprotein. The polyprotein had substantial amino acid sequence homology with those encoded by the RNAs of a Chinese isolate of sorghum mosaic virus (SrMV-C) and a Bulgarian isolate of maize dwarf mosaic virus, but it was most closely related to sugarcane mosaic virus (SCMV) isolates, for which only partial sequences have been published. According to the published criteria for distinguishing potyviruses, the sequence reported here is clearly a strain of SCMV, but it also showed a surprisingly high amino acid homology with SrMV-C in the HC-Pro, P3 and Cl proteins.     

The complete sequence of a sugarcane mosaic virus isolate causing maize dwarf mosaic disease in China

The complete sequence of a potyvirus from maize in Zhejiang Province was determined. The RNA was 9596 nucleotides long, excluding the 3'-poly (A) tail, and there was a single long open reading frame (ORF) of 9192 nts encoding a 346.1 ku polyprotein. The polyprotein had substantial amino acid sequence homology with those encoded by the RNAs of a Chinese isolate of sorghum mosaic virus (SrMV-C) and a Bulgarian isolate of maize dwarf mosaic virus, but it was most closely related to sugarcane mosaic virus (SCMV) isolates, for which only partial sequences have been published. According to the published criteria for distinguishing potyviruses, the sequence reported here is clearly a strain of SCMV, but it also showed a surprisingly high amino acid homology with SrMV-C in the HC-Pro, P3 and Cl proteins.