应用沁孢子和花药培养技术筛选油菜抗菌核病材料

油菜菌核病是世界油菜产区的重要病害之一 ,迄今尚未找到有效的控制方法。现有油菜品种中未见有能抗病的 ,双低油菜育种的成功诚然推动了油菜品质育种工作 ,但优质品种的抗菌核病能力下降。培育抗菌核病的优质油菜是世界性的研究难题。我们以探索创造油菜抗菌核病新种质的新途径为目的 ,开展了采用油菜小孢子 /花药培养 ,平阳霉素诱变和草酸胁迫相结合的方法 ,诱导和筛选抗病变异体的研究。取低芥酸和低硫甙油菜 (Ｂｒａｓｓｉｃａｎａｐｕｓ)品种“沪油 1 2”、品系“980 5”和“9841”的种子 ,表面灭菌后接种于诱导培养基 ( 2 /3ＭＳ Ｎ…     

油菜分子标记图谱构建及抗菌核病性状的QTL定位

以油菜品种０８５（抗菌核病）与湘油１３号杂交得到的Ｆ２代作为作图群体,以ＲＡＰＤ标记来构建油菜连锁图谱。通过对３００个１０－ｍｅｒ随机引物的筛选,共获得２００个多态性ＲＡＰＤ位点。经Ｍａｐｍａｋｅｒ／ＥＸＰ３．０软件处理,构建了１张含１９３个标记位点、１９个连锁群、覆盖长度为１３２４ｃＭ的连锁图谱。在此基础上利用Ｍａｐｍａｋｅｒ／ＱＴＬ １．１将抗菌核病基因定位在第四、八和十四３个连锁群上,即：Ｓ     

广西沿海受旱与咸酸田面积的分布与抗旱、耐盐种质资源鉴定

调查了广西沿海地区的北海、防城港、钦州3市8县(区)1975-2010年耕地面积、干旱受灾面积和咸酸田面积的动态变化以及抗旱、耐盐作物种类和分布;收集了一批水稻、木薯及芋头的地方种质资源,并对其进行了抗旱性和耐盐性的鉴定评价。本研究对广西沿海种质资源保护及利用,对抗旱、耐盐性研究及育种利用有重要意义。     

野生大豆资源对大豆疫病抗病性和耐病性鉴定

大豆疫病是大豆重要病害之一,在世界范围内导致严重经济损失。防治大豆疫病最有效方法是利用抗病或耐病品种。筛选抗性资源是发掘抗性基因和抗病育种的基础。本研究鉴定了野生大豆资源对大豆疫病的抗病性和耐病性,以期发掘优异抗源。苗期用子叶贴菌块方法鉴定104份野生大豆资源对两个不同毒力的大豆疫霉分离物PSJS2(毒力型:1a,1b,1c,1d,1k,2,3a,3b,3c,4,5,6,7,8)和PS41-1(毒力型:1a,1d,2,3b,3c,4,5,6,7,8)抗性,结果表明33份资源抗PS41-1,35份资源抗PSJS2,其中18份抗两个分离物。在抗病性鉴定基础性上,用菌层接种方法对选择的82份资源进行耐病性鉴定,发现7份高耐病性资源。这些结果表明,野生大豆中可能含有新的大豆疫病抗病和(或)耐病资源,这些抗病或耐病资源可以用于未来大豆抗病育种,以丰富大豆对大豆疫病的抗性遗传基础。     

植物抗旱耐盐基因的研究进展

近几年许多与植物抗旱耐盐相关基因被克隆和分析,同时通过转基因技术将这些基因转到植物中异源表达,能显著提高转基因植物的抗旱耐盐能力。这些基因主要包括渗透调节基因、蛋白类基因(如信号传导中的蛋白激酶基因)及转录因子等。在逆境条件下,渗透调节基因通过合成脯氨酸、甜菜碱、糖类和多胺类等渗透调节物质维持植物中的渗透平衡;蛋白激酶基因产物是细胞信号传导中的组分,这些基因能促进植物对干旱失水反应和逆境信号的传递,启动抗逆基因的表达;转录因子通过与相关基因的特异性结合来调控其表达,进而产生相关调控蛋白等物质增强植物在逆境中的生存能力。本文主要综述了这三类抗逆基因的研究现状及其生物学机理,讨论并分析这些基因在应用中尚待解决的问题,为发掘更多的抗逆性的基因资源和进一步开展分子育种工作提供参考。     

虉草营养器官解剖结构与抗旱耐涝性及利用之间的关系

为了从显微结构上进一步探讨虉草(Phalaris arundinacea L.)的抗旱耐涝性及与利用的关系,于2011年采用常规石蜡切片技术,对其根、茎叶3种营养器官进行解剖观察。结果表明,虉草根的结构自外而内依次为表皮、皮层、维管束鞘、初生韧皮部和初生木质部;茎由表皮、基本组织和维管束构成;叶片内部结构可分为表皮、叶肉和叶脉3部分。根皮层大的细胞间隙和气腔,初生木质部的后生大导管和茎基本组织解体形成的髓腔都是虉草良好的通气组织,是其耐水淹的主要显微特征。茎、叶片角质化的表皮和叶表皮所含的丰富泡状细胞组是虉草具有抗旱性的主要解剖结构特征。叶肉细胞排列紧密且只有少量气孔分布于叶片下表皮,这样的结构可减少蒸腾;叶肉细胞富含叶绿体,增强光合作用,获得更多的同化产物,确保了植株在干旱条件下也有足够的光合产物来维持正常的生理活动。茎、叶维管束部分大量的木纤维起到支撑作用。虉草根的皮层和维管柱部分、茎的基本组织和维管束部分、叶的叶脉部分都含有大面积的厚壁细胞,厚壁细胞中含有丰富的粗纤维和木质素。丰富的粗纤维、木质素等成分则是虉草能成为新能源燃料植物的必备条件。     

大黄酸抗耐甲氧西林金黄色葡萄球菌的抗菌机制

[背景]耐甲氧西林金黄色葡萄球菌(Methicillin-Resistant Staphylococcus aureus,MRSA)是医院及社区常见的机会性致病菌,具有多重耐药性、高发病率和高死亡率的特点.MRSA感染已成为全球医学界的普遍难题之一.[目的]研究大黄酸对MRSA的抗菌机制.[方法]以二倍稀释法测定大黄酸...     

抗菌豆豉发酵菌株的筛选及其脂肽组分鉴定和特性研究

目的:从豆豉中分离广谱抗菌活性菌株,并鉴定其发酵产物中的抗菌成分.方法:采用生理生化实验结合16SrRNA序列测定法鉴定目标分离株,通过LC-MS和ESI/CID技术对其抗菌组分进行鉴定.结果:从8个豆豉样品中分离出7株具有抗菌活性菌株,其中抗菌活性较强的分离株NT-6为枯草芽孢杆菌(Bacillus subtilis),其发酵液抗菌提取物的主要成分是抗菌脂肽类物质iturin、fengycin、surfactin同系物的混合物.该提取物对供试革兰氏阳性、阴性细菌、霉菌等指示菌具有广谱的抗菌活性,在pH 2～ 12、121℃条件下加热30min活性不丧失,说明具有广泛的pH适应范围和良好的耐热性.结论:该菌株产生多组分抗菌脂肽,其抑菌谱宽,适应性好,显示了在食品、农业、医药等领域具有良好的应用前景.     

小麦萌发期抗旱和耐盐性状的QTL分析

该研究以‘山农0431×鲁麦21’RIL群体及其父母本为材料,用20%PEG-6000溶液和100 mmol·L-1 NaCl溶液分别模拟干旱和盐环境,对12个小麦萌发期抗旱耐盐相关性状进行测定,结合已构建的分子标记遗传图谱对小麦萌发期抗旱、耐盐的相关性状进行QTL分析,为小麦抗旱、耐盐基因的克隆和分子标记辅助选择提供参考。结果表明:(1)正常、干旱和盐胁迫3种处理下共检测到143个QTL。检测到相对高频QTL(RHF-QTL)29个,平均贡献率范围为4.39%~13.28%,贡献率在10%以上的主效RHF-QTL有10个。(2)检测到胁迫下特异表达的RHF-QTL共17个,正常处理下特异表达的RHF-QTL为8个,稳定表达的RHF-QTL为4个。(3)QTL分析结果表明,7个RHF-QTL形成了3个QTL簇,且分布在2D、4D和5B等3条染色体上,其中:QC1位于2D染色体的wPt-6847~D-1172783区间,包括3个QTL(QRl-2D.2、QSdw-2D.3、QTdw-2D);QC2位于4D染色体短臂的D-2245724~D-1108531区间,包括2个QTL(QSl-4D、QShl-4D);QC3位于5B染色体的D-982263~S-1083095区间,包括2个QTL(QSl-5B.2、QTdw-5B.1)。     

一株新的耐辐射菌WGR702的分离鉴定及耐辐射特性

从辐射污染的土壤中分离到一株新的耐辐射菌WGR702.该菌为革兰氏阳性球菌,直径1.5 μm～2.5μm.菌体呈粉红色、能运动、兼性厌氧及不产孢子.其生长温度和pH范围分别为10℃～35℃和pH 5.0～10.0.(G C)mol%含量为60.5%.UV和γ射线辐射检测表明WGR702具有很强的耐辐射性.16S rDNA序列(EU315117)分析表明,菌株 WGR702与沙雷氏菌属(Serratia)菌株16S rDNA序列有很高相似性(94.79%～98.53%),但与沙雷氏菌属已报道菌株最大不同点是菌株WGR702细胞为球状,且革兰氏阳性.结合形态、生理生化特征分析表明菌株WGR702可能是沙雷氏菌属(Serratia)的一个新种.     

甘蓝型油菜与蔊菜的原生质体融合与植株再生

以甘蓝型油菜下胚轴和蔊菜叶片为外植体提取原生质体, 采用PEG-高pH、高Ca²⁺附加DMSO的原生质体融合方法, 用液体浅层静置培养融合体, 获得了10株融合杂种, 观察了杂种形态学和细胞学。结果表明：1%纤维素酶+0.2%离析酶+3 mmol/L MES 酶解14 h 可获得较高产率的油菜原生质体, 0.25% 纤维素酶+0.5%离析酶+5 mmol/L MES酶解12 h可获得较高产率的蔊菜原生质体; 30% PEG + 0.3 mol/L葡萄糖+50 mmol/L CaCl₂•2H₂O +15%DMSO的融合条件下, 获得了10.4%的融合率; 实验所获的原生质体融合材料可作为新种质。     

甘蓝型油菜与蔊菜属间杂种后代的苗期耐湿性综合评价

以生产上应用的耐湿性存在差异的3个甘蓝型油菜品种为对照,采用盆栽模拟湿害胁迫的方法,对15份源自甘蓝型油菜与蔊菜属间杂交获得的遗传稳定的创新种质进行苗期耐湿性鉴定和综合评价。结果显示:(1)湿害抑制根系生长,导致植株变矮,叶片发黄,生物量降低,湿害对根系影响最为严重,其次是植株总生物量积累和地上部分的生长。(2)通过主成分分析和隶属函数法,将全展叶数、绿叶率、苗高、根长、地上部分鲜重、根鲜重、植株总鲜重、地上部分干重、根干重和植株总干重等10个性状指标值转化成单一的综合评价值(D值),客观、科学、准确地对参试的18份材料进行了综合评价,为甘蓝型油菜苗期耐湿性评价提供了方法参考。(3)15份创新种质除绿叶率外的9个测定指标的耐湿性系数均高于对照品种,耐湿性综合评价结果显示,15份创新种质的D值均高于渝黄2号,13份创新种质的D值高于综合抗耐性俱佳的中油821,3份创新种质的D值高于高耐湿品种中双9号,聚类得到的高耐湿类群的7份材料中创新种质占6份(85.71%)。可见,创新种质的耐湿性平均要优于对照品种,说明通过远缘杂交获得的创新种质对甘蓝型油菜的耐湿性有了较为明显的改良和提高。     

Analysis of the Genetic Structure of Sclerotinia sclerotiorum (Lib.) de Bary Populations from Different Regions and Host Plants by Random Amplified Polymorphic DNA Markers

The genetic diversity and genetic structure of a population of isolates of Sclerotinia sclerotiorum (Lib.) de Bary from different regions and host plants were investigated using the random amplified polymorphic DNA (RAPD) method with 20 random decamer primer pairs in order to provide some information on the phylogenetic taxa and breeding for resistance to sclerotinia stem rot. A minimum of three and a maximum of 15 unambiguously amplified bands were generated, furnishing a total of 170 bands ranging in size from 100to 3 200 bp, corresponding to an average of 8.5 bands per primer pair. One hundred and four of these 170bands (61.2%) were polymorphic, the percentage of polymorphic bands for each primer pair ranging from 0.0% to 86.7%. The genetic relationships among the isolates, based on the results of RAPD analysis, were examined. The genetic similarity of all selected isolates was quite high. At the species level, the genetic diversity estimated by Nei's gene diversity (h) was 0.197 and S hannon's index of diversity (I) was 0.300. The unweighted pair-group mean analysis (UPGMA) cluster analysis showed that most isolates from the same regions were grouped in the same cluster or a close cluster. The population of isolates from Hefei (Anhui Province, China) was more uniform and relatively distant to other populations. The Canadian population collected from carrot (Daucus carota var. sativa DC.) was relatively close to the Polish population collected from oilseed rape (Brassica napus L.) plants. There was no relationship between isolates from the same host plants. An analysis of molecular variance (AMOVA) revealed that the percentage of variance attributable to variation among and within populations was 50.62% and 49.38%, respectively. When accessions from China, Europe, and Canada were treated as three separate groups, the variance components among groups,among populations within groups, and within populations were -0.96%, 51.48%, and 49.47%, respectively.The genetic differentiations among and within populations were highly significant (P ＜ 0.001). Similarly, the coefficient of gene differentiation (Gst) in total populations calculated by population genetic analysis was 0.229 4, which indicated that the genetic variation among populations was 22.94%. The gene flow (Nm)was 1.68, which indicated that the gene permutation and interaction among populations was relatively high.     

三种冬青属树种的耐涝性和耐旱性评价

通过致死性干旱和致死性水涝处理,用生理生态方法,对冬青(Ilexchinensis)、绿冬青(I.viridis)和无刺枸骨(I.cornatavar.fortunei)进行抗逆性研究。耐涝性结果表明随淹水时间延长,3种受淹冬青体内的游离脯氨酸和丙二醛含量增加,净光合速率下降;比较而言,绿冬青上述受淹反应出现早,无刺枸骨出现迟,而冬青介于二者之间;绿冬青耐涝约1周,无刺枸骨耐涝2周以上,冬青耐涝介于二者之间,在江南水乡推广利用,耐涝方面不会成为限制因素。耐旱结果表明随干旱的逐渐加重,3种冬青体内的游离脯氨酸含量呈上升趋势,比较而言,绿冬青上升的峰值出现早,冬青和无刺枸骨的上升峰值出现迟;绿冬青耐旱约15d,无刺枸骨耐旱约25d,冬青介于二者之间。3种冬青均有一定的抗逆性,其中无刺枸骨对水胁迫的适应能力最强,冬青次之,而绿冬青相对较弱。     

A new point mutation in the iron–sulfur subunit of succinate dehydrogenase confers resistance to boscalid in Sclerotinia sclerotiorum

Research has established that mutations in highly conserved amino acids of the succinate dehydrogenase (SDH) complex in various fungi confer SDH inhibitor (SDHI) resistance. For Sclerotinia sclerotiorum (Lib.) de Bary, a necrotrophic fungus with a broad host range and a worldwide distribution, boscalid resistance has been attributed to the mutation H132R in the highly conserved SdhD subunit protein of the SDH complex. In our previous study, however, only one point mutation, A11V in SdhB (GCA to GTA change in SdhB), was detected in S. sclerotiorum boscalid‐resistant (BR) mutants. In the current study, replacement of the SdhB gene in a boscalid‐sensitive (BS) S. sclerotiorum strain with the mutant SdhB gene conferred resistance. Compared with wild‐type strains, BR and GSM (SdhB gene in the wild‐type strain replaced by the mutant SdhB gene) mutants were more sensitive to osmotic stress, lacked the ability to produce sclerotia and exhibited lower expression of the pac1 gene. Importantly, the point mutation was not located in the highly conserved sequence of the iron–sulfur subunit of SDH. These results suggest that resistance based on non‐conserved vs. conserved protein domains differs in mechanism. In addition to increasing our understanding of boscalid resistance in S. sclerotiorum, the new information will be useful for the development of alternative antifungal drugs.     

Reaction of sunflower varieties to Sclerotinia sclerotiorum under various inoculation methods,time of field evaluation and phylogenic stages of host

Sunflower cultivation is affected seriously by Sclerotinia sclerotiorum (Lib.) de Bary in Iran, particularly north-western areas. Because of economic and environmental harms by chemical control, it is necessary to develop cultivars with adequate genetic resistance for reduction of yield losses. The purpose of this study was to find an effective method of inoculation with S. sclerotiorum under field evaluations. Three stem-inoculation techniques including: 1 – mycelium plug, 2 – oxalic acid solution (OAS) and 3 – infested wheat seeds with Sclerotinia mycelium were employed under field conditions. Four genotypes including Ghalami (local variety in market), Confeta, Allstar and Master were used in this study. The lesion length, lesion width and lesion as up and down leading on the stem from inoculation site were measured after 3, 7, 10 and 14 days of inoculation. The analysis of variance showed significant difference between all employed techniques and incubation days after inoculation. Mycelial plug (MP) inoculation technique produced significantly more developed lesions on the treated stems. In spite of this effect, Master variety demonstrated reasonable resistance reaction against the disease. The progress of disease in wounded treatments was also faster than the non-wounded ones. And, the shortest time to obtain significant differences between varieties was 10?days after inoculation. By comparison of results of lesion length at flowering and seed-filling stages, the more obvious effectiveness of the disease was observed at the second stage. Finally, there were negative correlations between mean temperature and mean lesion length in all three inoculation methods.     

ANGUSTIFOLIA negatively regulates resistance to Sclerotinia sclerotiorum via modulation of PTI and JA signalling pathways in Arabidopsis thaliana

Sclerotinia sclerotiorum is a devastating pathogen that infects a broad range of host plants. The mechanism underlying plant defence against fungal invasion is still not well characterized. Here, we report that ANGUSTIFOLIA (AN), a CtBP family member, plays a role in the defence against S. sclerotiorum attack. Arabidopsis an mutants exhibited stronger resistance to S. sclerotiorum at the early stage of infection than wild-type plants. Accordingly, an mutants exhibited stronger activation of pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) responses, including mitogen-activated protein kinase activation, reactive oxygen species accumulation, callose deposition, and the expression of PTI-responsive genes, upon treatment with PAMPs/microbe-associated molecular patterns. Moreover, Arabidopsis lines overexpressing AN were more susceptible to S. sclerotiorum and showed defective PTI responses. Our luminometry, bimolecular fluorescence complementation, coimmunoprecipitation, and in vitro pull-down assays indicate that AN interacts with allene oxide cyclases (AOC), essential enzymes involved in jasmonic acid (JA) biosynthesis, negatively regulating JA biosynthesis in response to S. sclerotiorum infection. This work reveals AN is a negative regulator of the AOC-mediated JA signalling pathway and PTI activation.     

Two consensus quantitative trait loci clusters controlling anthesis–silking interval, ear setting and grain yield might be related with drought tolerance in maize

Unravelling the molecular basis of drought tolerance will provide novel opportunities for improving crop yield under water-limited conditions. The present study was conducted to identify quantitative trait loci (QTLs) controlling anthesis–silking interval (ASI), ear setting percentage (ESP) and grain yield (GY). The mapping population included 234 F₂ plants derived from the cross X178 (drought tolerant) × B73 (drought susceptible). The corresponding F_2:3 progenies, along with their parents, were evaluated for the above-mentioned traits under both well-watered and water-stressed field conditions in three different trials carried out in central and southern China. Interval mapping and composite interval mapping identified 45 and 65 QTLs for the investigated traits, respectively. Two QTL clusters influencing ASI and ESP on chromosomes 1 (bin 1.03) and 9 (bins 9.03–9.05) were identified in more than two environments, showing sizeable additive effects and contribution to phenotypic variance; these two QTL clusters influenced GY only in one environment. No significant interaction was detected between the two genomic regions. A comparative analysis of these two QTL clusters with the QTLs controlling maize drought tolerance previously described in three mapping populations confirmed and extended their relevance for marker-assisted breeding to improve maize production under water-limited conditions.     

转基因技术在作物抗旱改良中的应用

以不同靶标基因为例,分不同作用机制（诸如渗透调节和清除氧自由基等）简要介绍了近10年来国内外利用转基因技术改良作物耐盐和抗旱性的研究进展,为我国北方农业的抗旱耐盐性研究提供一些思路。