红地球葡萄霜霉病系统型病株的田间分布型和抽样技术研究

为搞清红地球葡萄（Red Globe Grape）霜霉病（Plasmopara vaticola）的发生流行规律,以及在生产实践中为提高田间抽样的准确度,运用聚集度指数（K、CA、I＆和M^＊／X、λ）分析研究了红地球葡萄霜霉病系统型病株的田间分布型．结果表明：红地球葡萄霜霉病病株在葡萄园中的空间分布呈聚集型,分布的基本型是嵌纹分布和核心分布,适合率分别为75％和76．67％。二项分布的适合率为8．33％．在此分布型的基础上,通过调查病叶平均数,比较平行线、“W”字型、“Z”字型抽样方式,最后肯定平行线法最好．     

葡萄感霜霉病基因的分子标记(英文)

 在葡萄抗病育种中 ,幼苗期排除感霜霉病的后代具有特别重要的意义 .用 BSA,RAPD和SCAR方法研究了葡萄感霜霉病基因的分子标记 .分析了两个种间杂交组合 [毛葡萄 (抗病 )×欧洲葡萄 (感病 ) ]88- 1 1 0和 88- 84与 88- 1 1 0的 F1代自交或互交所得的 3个 F2 代 ,以及欧洲葡萄品种和中国野生葡萄种 .共筛选了 2 80个随机引物 .引物 OPO1 0产生了一个 RAPD标记 OPO1 0 - 80 0与葡萄感霜霉病主效基因紧密联锁 .将该 DNA片段克隆并测序 .OPO1 0 - 80 0的实际长度为 835bp,所以 OPO1 0 - 80 0应为 OPO1 0 - 835.据其两端序列 ,设计了一对长度为 2 6bp和 2 8bp的特异引物分别扩增上述试材 ,获得了与该 RAPD标记相同大小的一条带 ,将 RAPD标记转化为 SCAR标记SCO1 0 - 835.并证实了此 SCAR标记的通用性 ,该 SCAR标记可用于葡萄抗病育种中杂种后代对霜霉病的抗病与感病性鉴定 .     

高温诱导黄瓜抗霜霉病机理

研究了高温对黄瓜霜霉病菌致病力的影响以及高温控制霜霉病发生的效果.结果表明,40 ℃高温处理2 h和45 ℃处理1 h对黄瓜霜霉病的诱导抗病性作用最明显,其在接种后4 d时的防效分别为58.40%和45.81%,到接种后6 d时,防效分别下降为39.35%和37.65%.经高温诱导后,过氧化物酶（POD）、苯丙氨酸解氨酶(PAL)、几丁质酶（Cht）、β-1,3-葡聚糖酶（Glu）活性均显著高于对照,与未诱导植株相比,高温诱导后叶片组织的细胞壁表面有大量木质素沉积,表明高温处理后黄瓜表现出对霜霉病的抗病性.     

葡萄感霜霉病基因RAPD标记的序列分析

利用Ｗｉｚａｒｄ ＤＮＡ ｃｌｅａｎ－ｕｐ ｓｙｓｔｅｍ纯化葡萄感霜霉病基因ＲＡＰＤ遗传标记的ＤＮＡ片段,用细菌质粒ｐＧＥＭ Ｔ－ｅａｓｙ ｖｅｃｔｏｒ克隆该片段,采用自动荧光ＤＮＡ测序仪对片段的核苷酸组成进行双向测序。来自欧洲葡萄粉红玫瑰的葡萄感霜霉病基因ＲＡＰＤ标记由８３５对核苷酸及其特定序列组成。所获的感霜霉病基因ＲＡＰＤ标记可以作为合成探针的基础,用于葡萄抗病育种过程中的早期选择及品种对霜     

导入几丁酶基因育成抗葡萄白粉病的葡萄

日本《农业技术》,１９９９年５４卷３期第４Ｎ～５Ｎ页报道：葡萄以白粉病为主的、黑痘病、灰霉病、霜霉病等各种丝状菌病,是葡萄栽培的大问题。对此,日本果树试验场的山本等把取自水稻的几丁酶基因导入葡萄,育成抗葡萄白粉病的葡萄。方法是从葡萄品种‘新玫瑰香’的培养细胞育成不定胚,与具有水稻几丁酶基因（ＲＣＣ２）的土壤杆菌共培养,用抗菌素选择得到的重组不定胚再分化成植物体,将几丁酶基因导入葡萄,对获得的许多再生植物体驯化、育成。首先,用ＰＣＲ法确认已导入基因,从中选择了８个性状转化葡萄个体作扦插苗,将约１０…     

不同品种葡萄抗霜霉病特性与叶片POD、PPO活性关系的研究

在霜霉病盛发期,对8804、梅尔诺、品丽珠3个葡萄品种(系)叶片中的PPO和POD活性变化进行了测定.结果显示,8804的PPO和POD活性较大,并保持相当长时间的高活性值,而梅尔诺、品丽珠叶片中PPO和POD活性较小;8804的PPO酶活性变化范围高于其它2个品种,但POD酶活性变化范围却低于后者.葡萄叶片中PPO和POD活性与葡萄霜霉病抗性之间存在一定的相关性,且不同抗感品种间PPO和POD酶活性存在极显著差异.研究结果表明,8804较梅尔诺、品丽珠对霜霉病具有较强的抗性.     

美国酿酒葡萄品种在北京地区的生长和适应性表现

调查研究了首次引进的13个美国酿酒葡萄品种在北京地区栽培后的植物学性状、果实品质和抗寒及抗霜霉病能力,并与传统的欧亚种酿酒葡萄及砧木品种进行了比较与分析,总结了它们之间的植物学性状与抗逆性的差异。本试验选出了在抗寒及抗霜霉病方面表现优良的美国酿酒葡萄品种,为这些品种在我国的实际栽培推广提供科学依据。     

山葡萄(Vitis amurensis)资源核心种质的初步构建

采用省份分组-花型分组-聚类-分组法构建了山葡萄核心种质,在此基础上,考虑到山葡萄品种在生产上和雄株山葡萄在科学研究上的作用,把未进入核心种质的山葡萄品种和雄株山葡萄也并入核心种质。该核心种质包括48份资源（占资源总数的31％）,其中来自吉林省的资源18份（占37．5％）,黑龙江省的资源29份（占60．4％）,辽宁省的资源1份（占2．1％）;两性花资源22份（占45．8％）,雌能花资源21份（占43．8％）,雄株5份（占10．4％）。除出汁率外,其他性状的符合度达72％以上;除果粒质量外,其他性状的变幅吻合度达79％以上。以上结果表明,所构建的核心种质较好地代表了评价的山葡萄资源。     

钙肥对葡萄根瘤蚜生长发育和繁殖的影响

【目的】为明确葡萄所需的大量元素钙对葡萄根瘤蚜 Daktulosphaira vitifoliae(Fitch）生长发育和繁殖的影响。【方法】在温室中,设置不同 Ca⁽²⁺⁾浓度（0、2 和 4mmol·L(2+)浓度（0、2 和 4mmol·L^(-1)）的钙全营养液于葡萄全生育期浇灌葡萄,评价离体根段对葡萄根瘤蚜的存活率、若蚜、成蚜寿命和繁殖力的影响。【结果】随着钙离子浓度的增加,葡萄根瘤蚜在葡萄根上的 1 龄若蚜历期缩短 0.80 倍以上,成蚜历期增加超过 1.88倍,寿命显著延长超过 1.18 倍,产卵量提高 3.34 倍以上。与 0 mmol·L(-1)）的钙全营养液于葡萄全生育期浇灌葡萄,评价离体根段对葡萄根瘤蚜的存活率、若蚜、成蚜寿命和繁殖力的影响。【结果】随着钙离子浓度的增加,葡萄根瘤蚜在葡萄根上的 1 龄若蚜历期缩短 0.80 倍以上,成蚜历期增加超过 1.88倍,寿命显著延长超过 1.18 倍,产卵量提高 3.34 倍以上。与 0 mmol·L^(-1) Ca(-1) Ca⁽²⁺⁾和 2 mmol·L(2+)和 2 mmol·L^(-1) Ca(-1) Ca⁽²⁺⁾处理相比,葡萄根瘤蚜在 4 mmol·L(2+)处理相比,葡萄根瘤蚜在 4 mmol·L^(-1) Ca(-1) Ca⁽²⁺⁾处理下的葡萄根上,内禀增长率 r_m、周限增长率 λ 和净增殖率 R_0均最高,分别为（0.191 ± 0.003）、（1.210 ± 0.004）和（169.070 ± 11.897)(P < 0.05）,且世代发育历期显著缩短（P <0.05）。特定年龄龄期存活率 S_(xj)、种群特定年龄繁殖力 m_x和种群特定年龄繁殖值 l_xm_x在 4mmol·L(2+)处理下的葡萄根上,内禀增长率 r_m、周限增长率 λ 和净增殖率 R_0均最高,分别为（0.191 ± 0.003）、（1.210 ± 0.004）和（169.070 ± 11.897)(P < 0.05）,且世代发育历期显著缩短（P <0.05）。特定年龄龄期存活率 S_(xj)、种群特定年龄繁殖力 m_x和种群特定年龄繁殖值 l_xm_x在 4mmol·L^(-1)Ca(-1)Ca⁽²⁺⁾处理种群中保持较高水平,随着钙浓度降低,葡萄根瘤蚜的各生命参数明显降低。【结论】钙元素对葡萄根瘤蚜的生长发育和繁殖能力具有促进作用,葡萄根瘤蚜发生葡萄园更应合理使用钙肥。     

58种植物提取液对葡萄霜霉病菌的抑菌活性筛选研究

通过 5 8种植物提取液对葡萄霜霉病菌 (Plasmoparaviticola)的游动孢子囊萌发抑制率及接种叶圆盘的发病控制效果的研究 ,表明 :虎耳草 (ChlorisvirgataSwartz) ,黄檀 (DalbergiahuoeanaHance) ,马尾松 (PinusmassonianaLarmb .) ,牡丹 (Paeoniasuffruticosa) ,刺槐 (RobiniapseudoacciaL .) 5种植物提取液对葡萄霜霉病有抑菌效果 ,对游动孢子囊萌发抑制率分别为 :84 70 %、87 5 2 %、88 4 6 %、83 78%、83 15 % ,而叶圆盘生测结果 ,其发病率均为 0级 ,该效果和目前常用药剂波尔多液的效果等同 ,有的还优于对照。该研究为开发植物源控制葡萄霜霉病杀菌剂奠定了基础。     

黄瓜霜霉病抗病基因的RAPD及SCAR标记

以感霜霉病黄瓜L18-10-2和抗霜霉病黄瓜129为亲本构建F2代分离群体,以F3代植株霜霉病抗性鉴定表示F2代各单株抗病性并得以区分各单株杂合或纯合感病性,采用RAPD技术和转SCAR的方法筛选黄瓜抗霜霉病基因分子标记.结果显示,在318条RAPD引物中有18条引物表现出两亲本间多态性,其中引物P18的SB-SP18561扩增片段与霜霉病抗病基因之间紧密连锁,根据交换率和Kosambi函数公式计算其遗传距离为7.85 cM.回收SBSP18561片段并克隆和测序,其准确长度为561 bp.将该RAPD标记转换为SCAR标记,长度为494 bp,命名为SSBSP18494.     

A Basic Peroxidase Isoenzyme, Marker of Resistance Against Plasmopara viticola in Grapevines, is Induced by an Elicitor from Trichoderma viride in Susceptible Grapevines

The constitutive expression of basic peroxidase isoenzymes in the Plasmopara viticola -resistant ( Vitis vinifera × Vitis rupestris) × Vitis riparia crossing and in the P. viticola -susceptible V. vinifera parent species was studied. The results illustrate that both leaves and stems of the ( V. vinifera × V. rupestris) × V. riparia crossing showed the differential expression of a basic peroxidase isoenzyme B₃ (pl = 8.9), this being almost completely absent from the P. viticola -susceptible V. vinifera parent species. To test whether the basic peroxidase isoenzyme B₃ may be considered as a molecular marker of disease resistance in Vitis spp., suspension cell cultures derived from the P. viticola -susceptible V. vinifera parent species were treated with an elicitor (cellulase Onoztika R-10) from the soil fungus Trichoderma viride , a specific and well-known elicitor of disease resistance reactions in grapevines. The results showed that treatment with the elicitor induces, simultaneously with the activation of the disease resistance mechanism, the appearance of B₃ in the cell cultures. These results suggest that the basic peroxidase isoenzyme B₃ may be considered as a marker of disease resistance in Vitis species since it is present in the P. viticola -resistant ( V. vinifera × V. rupestris) × V. riparia hybrid and is induced by the elicitor Onozuka R-10 in cell cultures of the P. viticola -susceptible Vitis vinifera parent species. This conclusion is supported by the presence of this isoenzyme in other resistant and its absence in other susceptible Vitis spp.     

SCAR markers for discriminating species of two genera of medicinal plants, Liriope and Ophiopogon

The development of DNA markers that can closely discriminate between Liriope and Ophiopogon species is vital for efficient and accurate identification of these species, and to ensure the quality, safety, and efficacy of medicines made from these plants. We developed species-specific molecular markers for these two genera. Forty RAPD primers were tested to detect polymorphism; species-specific RAPD bands were gel-purified, cloned, and sequenced. Primers for sequence-characterized amplified regions (SCARs) were then designed, based on nucleotide sequences of specific RAPD primers. SCAR markers SA06 and SB05, specific to Ophiopogon japonicus, amplified 460- and 553-bp DNA fragments, respectively. The marker SA12 amplified a 485-bp fragment specific to Liriope platyphylla. This is the first report of a species-specific SCAR marker for this group. These markers will be useful for rapid identification of closely related Liriope and Ophiopogon species.     

Association of RGA-SSCP markers with resistance to downy mildew and anthracnose in grapevines

Downy mildew (Plasmopara viticola) and anthracnose (Sphaceloma ampelinum) are two major diseases that severely affect most grapevine (Vitis vinifera) cultivars grown commercially in Thailand. Progress of conventional breeding programs of grapevine for improved resistance to these diseases can be speeded up by selection of molecular markers associated with resistance traits. We evaluated the association between 13 resistance gene analog (RGA)-single-strand conformation polymorphism (SSCP) markers with resistance to downy mildew and anthracnose in 71 segregating progenies of seven cross combinations between susceptible cultivars and resistant lines. F(1) hybrids from each cross were assessed for resistance to downy mildew and anthracnose (isolates Nk4-1 and Rc2-1) under laboratory conditions. Association of resistance traits with RGA-SSCP markers was evaluated using simple linear regression analysis. Three RGA-SSCP markers were found to be significantly correlated with anthracnose resistance, whereas significant correlation with downy mildew resistance was observed for only one RGA-SSCP marker. These results demonstrate the usefulness of RGA-SSCP markers. Four candidate markers with significant associations to resistance to these two major diseases of grapevine were identified. However, these putative associations between markers and resistance need to be verified with larger segregating populations before they can be used for marker-assisted selection.     

RAPD-based SCAR marker SCA 12 linked to recessive gene conferring resistance to anthracnose in sorghum [Sorghum bicolor (L.) Moench]

Anthracnose, caused by Colletotrichum graminicola, infects all aerial parts of sorghum, Sorghum bicolor (L.) Moench, plants and causes loss of as much as 70%. F₁ and F₂ plants inoculated with local isolates of C. graminicola indicated that resistance to anthracnose in sorghum accession G 73 segregated as a recessive trait in a cross with susceptible cultivar HC 136. To facilitate the use of marker-assisted selection in sorghum breeding programs, a PCR-based specific sequence characterized amplified region (SCAR) marker was developed. A total of 29 resistant and 20 susceptible recombinant inbred lines (RILs) derived from a HC 136 × G 73 cross was used for bulked segregant analysis to identify a RAPD marker closely linked to a gene for resistance to anthracnose. The polymorphism between the parents HC 136 and G 73 was evaluated using 84 random sequence decamer primers. Among these, only 24 primers generated polymorphism. On bulked segregant analysis, primer OPA 12 amplified a unique band of 383 bp only in the resistant parent G 73 and resistant bulk. Segregation analysis of individual RILs showed the marker OPA 12₃₈₃ was 6.03 cM from the locus governing resistance to anthracnose. The marker OPA 12₃₈₃ was cloned and sequenced. Based on the sequence of cloned RAPD product, a pair of SCAR markers SCA 12-1 and SCA 12-2 was designed using the MacVector program, which specifically amplified this RAPD fragment in resistant parent G 73, resistant bulk and respective RILs. Therefore, it was confirmed that SCAR marker SCA 12 is at the same locus as RAPD marker OPA 12₃₈₃ and hence, is linked to the gene for resistance to anthracnose.     

油菜单显性核雄性不育基因的分子标记

以陕西省杂交油菜研究中心选育的单显性核不育油菜分离群体为材料,利用集群分离法（BSA）对该油菜单显性核不育基因进行了RAPD分析。在随机选取的300个10碱基随机引物中,引物S243（5′CTATGCCGAC3′）在可育集团与不育集团间扩增出特异而可重复的1．5kb的多态性片段OPU-031500,而在细胞质雄性不育和其它核不育类型油菜中均未扩增出上述特异性片段,从而确证此RAPD标记OPU-031500。片段是与甘蓝型油菜单显性核不育基因连锁的。将该多态性片段克隆并测序,发现其序列与拟南芥的一段DNA序列高度同源。根据同源序列及测序结果设计两对特异引物（P1／P2和P3／P4）,引物P3／P4在可育系中可扩增到约1．5kb的单一特异片断,而在不育系中无带,从而将RAPD标记转化为稳定可靠的SCAR标记。     

西瓜抗枯萎病育种分子标辅助选择的研究

将西瓜野生种质ＰＩ２９６３４１抗枯萎病生理小种１的抗性基因连锁的ＲＡＰＤ标记ＯＰＰ０１．７００进行克隆、测序,Ｓｏｕｔｈｅｒｎ杂交证明此标记为１个单拷贝,并转化为ＳＣＡＲ标记,简化了ＳＣＡＲ扩增产物的检测技术。上述技术在抗病转育后代造反中得到了很好的应用,初步建立了西瓜抗枯萎病育种分子标记辅助选择技术系统。