植物内生菌在生物防治中的应用

从植物内生菌的抗病虫机理及生防作用方面综述了植物内生菌在生物防治中的研究进展,对植物内生菌的抗病虫机理进行了探讨,并展望了植物内生菌在植物保护中的应用前景.     

抑制消减杂交法分离紫花苜蓿幼苗铝胁迫诱导表达的cDNA

利用抑制消减杂交(SSH)技术分离铝胁迫诱导紫花苜蓿差异表达的基因,以水培试验获取的中苜一号幼苗为材料,以80μmol/L铝离子胁迫的紫花苜蓿作为试验组,未胁迫的为驱动组,构建了一个包含456个克隆的SSH文库。对构建的文库进行鉴定,随机选取20个阳性克隆测序,共获得15条有效EST序列,然后将测序结果提交到GenBank进行Blastn比对,获得了3条未知基因的序列,推测它们可能与植物的抗铝作用有关。检测结果表明,构建的文库质量较好,可以进行进一步深入研究,为揭示植物耐铝性的分子机理提供理论基础。     

植物基因cDNA克隆新技术及进展

过去人们对植物基因的cDNA克隆主要集中在植物看家基因 (house keepinggene) ,而对植物发育调控基因 (developmentalregulatedgene)进行的cDNA克隆研究工作开展得比较少。随着近年来基因差异表达研究技术的飞速发展 ,已建立了好几种发育调控基因的cDNA克隆方法 ,如从较早的DD PCR到最近的cDNARDA、SSH和IPGI,着重论述了cDNARDA、SSH和IPGI的基本技术策略 ,并对三者之间进行了比较分析。     

植物抗盐机理研究进展

对近年来植物抗盐机理的研究进展作了概述,阐明了植物对盐分的反应及盐分对植物的不同伤害,并从盐生植物的形态、生理和分子水平上综述了盐生植物的抗盐机理,最后对今后植物抗盐机理的研究可能存在的问题提出了自己的观点.     

植物基因工程的应用与研究进展及潜在风险性分析

简要综述近年来植物基因工程在改善植物性状,提高植物抗性等遗传育种方面的应用与研究进展。重点介绍了植物基因工程的应用,如改良农产品品质,提高农作物抗病虫毒、抗除草剂、抗自然灾害的能力,开发疫苗等。分析了植物基因工程在农作物育种方面的应用前景及其潜在的风险性。     

植物引种驯化过程中的病虫害问题及其防治对策

植物园在引种驯化过程中存在的病虫害问题是：病虫随植物的引种大量带入园区,且严重为害,造成很大损失;生境的改变及栽培管理的不当使植物的抗病虫能力下降;农药的过分使用,造成环境污染,病虫抗药性的产生及对有益生物的大量杀伤,破坏了自然界的生态平衡。防治对策提出了加强植物检疫及栽培管理、利用生物防治、合理使用化学农药等措施。     

激活蛋白处理水稻引发基因差异表达的研究

利用抑制性消减杂交技术(Suppression Subtractive Hybridization,SSH)成功构建了植物激活蛋白处理水稻与非处理组水稻中差异表达的消减cDNA文库。从文库中一共筛选到1756个克隆,通过反向Northern杂交,从中得到264个有效克隆并测序,利用BLAST在GenBank数据库进行序列相似性比对分析,获得28个上升表达差异基因。通过分析这些基因与植物的光合作用、营养物质的运输代谢等多种植物的生理生化功能相关。本研究为揭示激活蛋白的作用机理奠定基础。     

抑制差减杂交(SSH)技术及其在植物基因分离上的应用

SSH是一种基于抑制PCR和差减杂交技术建立的 ,在转录水平上研究基因表达的技术 ,具有稳定、高效、可靠的特点 ,可对生物的生长、发育、衰老、死亡等生命过程及生物或非生物逆境胁迫对生物所造成的影响等进行全面、系统的分析。简单介绍了SSH的基本原理、技术要点 ,并对技术本身的改进和提高及在转录水平上与其它技术方法的比较及其在植物基因分离上的应用进行了概述。     

抗病原菌植物基因工程进展

植物病原菌给农林生产带来巨大的损失,植物基因工程在培育抗病原菌植物方面是传统育种技术的补充和发展,短短几年,在抗细菌和抗真菌植物基因工程方面出现了一些全新的成功策略,这些范例都是针对病原菌的生理结构、致病机理及与植物的相互关系。本文概括论述了这些策略的基本思路并对其局限性加以探讨。随着植物病理学、植物分子生物学和病原菌分子生物学的研究进展,新的抗性策略将会出现。     

抑制消减杂交技术(SSH)及其在植物基因克隆上的应用

介绍了抑制消减杂交技术（SSH）的主要原理、基本方法、优越性及主要缺陷,并就其在植物基因克隆上的应用作一综述。     

Transcriptome analysis of sugar beet root maggot (Tetanops myopaeformis) genes modulated by the Beta vulgaris host

Sugar beet root maggot (SBRM, Tetanops myopaeformis von Röder) is a major but poorly understood insect pest of sugar beet (Beta vulgaris L.). The molecular mechanisms underlying plant defense responses are well documented, however, little information is available about complementary mechanisms for insect adaptive responses to overcome host resistance. To date, no studies have been published on SBRM gene expression profiling. Suppressive subtractive hybridization (SSH) generated more than 300 SBRM ESTs differentially expressed in the interaction of the pest with a moderately resistant (F1016) and a susceptible (F1010) sugar beet line. Blast2GO v. 3.2 search indicated that over 40% of the differentially expressed genes had known functions, primarily driven by fruit fly D. melanogaster genes. Expression patterns of 18 selected EST clones were confirmed by RT‐PCR analysis. Gene Ontology (GO) analysis predicted a dominance of metabolic and catalytic genes involved in the interaction of SBRM with its host. SBRM genes functioning during development, regulation, cellular process, signaling and under stress conditions were annotated. SBRM genes that were common or unique in response to resistant or susceptible interactions with the host were identified and their possible roles in insect responses to the host are discussed.     

植物-昆虫间的化学通讯及其行为控制

在植物与昆虫间的化学通讯中植物气味物质起着决定性的作用,它调控着昆虫的多种行为,诸如引诱昆虫趋向寄主植物,刺激昆虫取食,引导昆虫选择产卵场所,进行传粉和防御昆虫等。有些植物则当受到食植性昆虫危害时会释出一些引诱害虫天敌的化学信号。这些化学信号是一些挥发性萜类混合物,天敌昆虫就以此来区分受害和未受害植株。尽管目前在害虫综合治理中,昆虫信息素的应用越来越显得比天然植物气味源更受重视,但是必须指出的是,昆虫信息化合物首次成功地使用于植物保护的却是天然植物气味源。在利用植物气味源作害虫测报和防治中,近年来一种简单价廉的粘胶诱捕器己成为多种害虫的标准测报工具。在害虫综合治理中利用植物气味源的技术显然是具有不可估量的潜力。文中提出了利用基因工程技术来改造植物,使植物能释放特定的驱避剂或其它控制昆虫行为的特殊气味物质的新概念。     

番茄茎腺毛差异表达序列分离与分析

腺毛是参与植物防御性反应与次生代谢挥发物合成的特异化器官,了解腺毛专一性表达序列标签将有助于我们进一步认识植物腺毛的特异性功能.本文应用磁珠介导的抑制消减杂交方法(SSH),以番茄茎腺毛为检测子(tester),去除腺毛的番茄茎为驱动子(driver),构建番茄茎腺毛差异表达cDNA文库,分离腺毛差异表达基因.随机挑选了108个差异ESTs进行测序,测序结果使用Blast2go程序进行blastx比对、功能注释和KEGG代谢路径分析.结果表明,绝大部分ESTs功能与胁迫响应、物质代谢、生物及非生物刺激反应相关,具有结合、催化功能.这些分离的差异ESTs为进一步研究番茄腺毛的植物防御性机制奠定了基础.     

Isolation and Analysis of Differential Expressed ESTs from Stem Trichomes of Lycopersicon esculentum (Solanaceae)

Glandular trichomes are special organs involved in plant defense response and synthesis of volatile secondary metabolites, analyzing trichome specific expressed sequence tags will help us further understand the specific function of plant trichomes. In this paper, suppression subtractive hybridization（SSH） based on magnetic beads technology was used to isolate differential expressed genes of the glandular trichomes in Lycopersicon esculentum. The differential expressing cDNA library was constructed using the glandular trichomes cDNA as tester and the cDNA from the stem without glandular trichomes as driver. After randomly sequencing 108 differential ESTs, Blast2go program was used to do blastx, functional annotation and metabolism analysis. The results show that most ESTs are related to substance metabolism, response to stress, biotic or abiotic stimulus, and have binding and catalytic function. These differential genes lay the foundation for further research on defense mechanism of the tomato trichomes.     

Construction of suppression subtractive hybridization libraries and identification of brown planthopper-induced genes

A suppression subtractive hybridization technique was used to screen for brown planthopper (BPH)-inducible genes in rice (Oryza sativa). cDNAs from a BPH-resistant rice line (B5) infested by BPH were used as the tester population, and mixed cDNAs from a BPH-sensitive line (MH63) and a control (uninfested B5) as the driver population. After hybridizing and cloning, forward and reverse subtraction cDNA libraries were obtained, containing 5700 clones. These clones were further analyzed by differential gene expression screening, and 154 clones that were clearly induced by BPH were identified. Sequencing analysis and homology searching showed that these clones represent 136 single genes, which were assigned to functional categories, including 10 putative cellular functions, according to categories established for Arabidopsis. The 136 genes include 21 known to be related to disease, wound and other stresses, most of which were found to be up-regulated in BPH feeding responses. In addition, an Oryza cysteine inhibitor and a beta-glucosidase belonging to the 21 genes group were found in the rice response to BPH feeding, these two genes have previously been shown to be induced in plant responses to chewing insects. Our results not only confirm that several identical genes are activated in defense mechanisms against both sucking and chewing insects, but also show that genes have overlapping functions in both pathogen and insect resistance.     

耐药淋球菌相关核苷酸序列的克隆与初步分析*

为克隆淋球菌染色体耐药相关核苷酸序列,我们利用抑制性消减杂交技术构建耐药性淋球菌与标准参考菌株差异DNA消减库,从中筛选淋球菌耐药相关核苷酸序列。通过初步筛选,对克隆得到的DNA片段进行测序,经GenBank和淋球菌基因组序列库检索分析,发现5个未知新核苷酸序列。这些核苷酸序列可能与淋球菌染色体耐药性相关,将为研究淋球菌耐药性提供新的实验对象。     

Rice–gall midge interactions: Battle for survival

Gall midges are insects specialized in maneuvering plant growth, metabolic and defense pathways for their benefit. The Asian rice gall midge and rice share such an intimate relationship that there is a constant battle for survival by either partner. Diverse responses by the rice host against the midge include necrotic hypersensitive resistance reaction, non-hypersensitive resistance reaction and gall-forming compatible interaction. Genetic studies have revealed that major R (resistance) genes confer resistance to gall midge in rice. Eleven gall midge R genes have been characterized so far in different rice varieties in India. In addition, no single R gene confers resistance against all the seven biotypes of the Asian rice gall midge, and none of the biotypes is virulent against all the resistance genes. Further, the interaction of the plant resistance gene with the insect avirulence gene is on a gene-for-gene basis. Our recent investigations involving suppressive subtraction hybridization cDNA libraries, microarray analyses, gene expression assays and metabolic profiling have revealed several molecular mechanisms, metabolite markers and pathways that are induced, down-regulated or altered in the rice host during incompatible or compatible interactions with the pest. This is also true for some of the pathways studied in the gall midge. Next generation sequencing technology, gene expression studies and conventional screening of gall midge cDNA libraries highlighted molecular approaches adopted by the insect to feed, survive and reproduce. This constant struggle by the midge to overcome the host defenses and the host to resist the pest has provided us with an opportunity to observe this battle for survival at the molecular level.