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几丁质酶(Chitinase)基因转化小麦的研究

本研究利用基因枪法、农杆菌法和花粉管通道法,首次将菜豆几丁质酶基因导入春小麦东农7742中,同时对转基因的表达和遗传进行了分析,对转基因植株的抗病性进行了观察。对影响转化的各种因素和转基因小麦研究中存在的问题提出了自己的见解。主要研究结果如下:1.利用PDS1000/He型高压氦气基因枪,以携带菜豆几丁质酶基因的质粒DNA轰击小麦萌动幼胚8139块,在东农7742To代获得经PCR扩增和PCR-Southern杂交证实的转基因小麦植株7株,转基因植株分属4个不同轰击参数的组合,转化率分别为0.56%、0.36%、0.20%和0.35%;利用农杆菌介导法和利用基因枪与农杆菌协同转化法转化东农7742小麦幼胚愈伤,共感染外植体1546块,获得转化愈伤组织的频率分别为0.46%和0.32%,但转化愈伤未能再生植株;利用花粉管通道法将菜豆几丁质酶基因导入小麦东农7742,共操作小花385个,获得2株转化植株,转化率为0.52%;三种转化  (本文共89页) 本文目录 | 阅读全文>>

四川农业大学
四川农业大学

小麦成熟胚受体筛选及基因枪法转几丁质酶基因植株获得和插入位点研究

小麦是世界上最重要的农作物之一,也是我国种植面积最广的主要农作物之一,小麦的稳产高产直接关系着我国的经济发展和国民生活水平的提高。然而由于自然灾害、病虫害等因素的影响,使得小麦的产量大幅度下降。因此,努力提高小麦的产量和质量是现在小麦育种所面临的首要问题。基因工程育种克服了传统的育种方法育种周期长等的弱点,为小麦育种提供了一条更新的途径。通过基因工程育种,将抗虫、抗病、抗逆、增产等基因转入小麦中已经成为小麦育种的主要目的。另外,直接对农艺性状优良的小麦品种进行转基因研究会更快速的得到具有应用价值的小麦新品种。因此,筛选出具有优良组织培养特性的高产小麦品种,对小麦基因工程的发展有着重要的意义。本研究选择7个小麦品种绵阳11、川农20、川农21、川农23、川农24、川农25、川农26为实验材料,剥取小麦成熟胚并将胚根切除后进行组织培养,通过观察其组织培养特性筛选出了绵阳11和川农23这两个具有优良组培特性并且农艺性状较好的小麦基因型...  (本文共61页) 本文目录 | 阅读全文>>

《Journal of Central South University of Technology》2009年04期
Journal of Central South University of Technology

Purification and characterization of extracellular chitinase from a novel strain Aspergillus fumigatus CS-01

1 Introduction Chitin, a linear polymer of β-1, 4-N-acetylglucose- amine units with abundance next to cellulose, is the most abundant natural amino polysaccharide [1]. It is widely distributed as a structural component of crustaceans, insects, as well as a component of the cell walls of most fungi and some algae [2]. Previous reports indicated that these natural polymers and their degraded products have excellent pro...  (本文共6页) 阅读全文>>

《浙江大学学报(农业与生命科学版)》2004年04期
浙江大学学报(农业与生命科学版)

Study on the character of chitinase produced by Trichoderma spp. with measuring reducing sugar

A trusty and intuitionistic method for screening chitinase produced by Trichoderma spp. was developed. 38 isolates of Trichoderma spp. were cultured in liquid medium with chitin or colloidal chitin as the sole carbon source for 4 days. The supernatant of the fermented broth ...  (本文共1页) 阅读全文>>

《浙江大学学报(农业与生命科学版)》2004年04期
浙江大学学报(农业与生命科学版)

Are mycoparasitism and chitinase production species or isolate dependent in Trichoderma?

The relationship between taxonomic status of Trichoderma spp., chitinase production in solid substrate fermentation (SSF) on four media and mycoparasitism in dual culture (confrontation assay) against four plant pathogenic fungi was studied. Seventy five Trichoderma isolates belonging to 35 species have been screened. The plant pathogenic fungi used in confrontation assay were Botrytis cinerea, Fusarium oxysporu...  (本文共1页) 阅读全文>>

内蒙古大学
内蒙古大学

细菌表达dsRNA和寄主表达hairpin dsRNA介导的玉米黏虫chitinase基因的RNA干扰研究

玉米黏虫(Mythimna separata)属于鳞翅目夜蛾科,是一种重要的农业害虫,严重暴发时会造成农作物大面积减产甚至绝收。RNA干扰(RNA interference,RNAi)是指生物体内源或者外源的双链RNA(double-stranded RNA,dsRNA)介导产生特异性的转录后基因沉默的现象。RNAi除了在模式昆虫如果绳(D.melannogaster)中得到广泛应用以外,在非模式昆虫中也得到了成功的应用。进一步研究发现,RNA干扰技术与第二代测序技术相结合,可以快速找到非模式昆虫的致死效应的靶基因序列,加快了利用RNAi技术控制害虫的步伐。本研究以玉米黏虫几丁质酶编码基因(chitinase)为候选基因,分别设计合成了chitinase 和chitinase2基因RNA干扰序列,为了达到RNA干扰效应,我们设计了两个实验方案。第一,采用了细菌表达dsRNA的RNA干扰系统(即大肠杆菌HT115和L4440载体)...  (本文共64页) 本文目录 | 阅读全文>>

《Agricultural Sciences in China》2008年12期
Agricultural Sciences in China

Purification and Characterization of a Novel Thermostable Chitinase from Thermomyces lanuginosus SY2 and Cloning of Its Encoding Gene

INTRODUCTION Chitin, the second most abundant natural biopolymer after cellulose, consists of a linear β-1,4-homopolymer of N- acetylglucosamine residues (Haki and Rakshit 2003). The covering layer of insects, cell walls of various fungi and crab, and shrimp wastes are the main sources of chitin (Majeti and Kumar 2000). The basic parts of shrimp and prawns used for consumption are the muscle tissues. Thus, the waste ...  (本文共8页) 阅读全文>>