T-DNA插入产生的水稻白化苗突变的遗传分析

在筛选和鉴定水稻T-DNA(含Basta抗性基因Bar)插入纯合体的过程中,观察到1个白化苗的突变体.对该突变体进行遗传分析表明,分离群体出现绿苗和白化苗2种类型,其分离比率为3:1,符合1对基因的显性遗传.Basta 抗性检测、PCR分子检测及Southern杂交证实,该突变体是由单一T-DNA插入所引起的,白化苗性状与T-DNA共分离.该突变材料可用于插入座位的基因克隆.     

T-DNA (Ds) 插入产生的水稻卷叶突变的遗传分析

在筛选和鉴定水稻T-DNA（Ds）插入纯合体的过程中,观察到1个叶片卷曲的突变体．对该突变体进行遗传分析表明,分离群体出现叶片卷曲和叶片正常2种植株类型,其分离比率为3：1,符合1对基因的显性遗传．Basta抗性检测及PCR分子检测证实,该突变体是由单一T-DNA（Ds）插入所引起的,突变性状与T-DNA（Ds）共分离．该突变材料可用于插入座位的基因克隆．     

水稻T-DNA插入突变体库筛选及其突变特征研究

采用水培方法大规模苗期筛选水稻T-DNA插入突变体库,以苗期表观症状为参考指标,从2 173个T1代家系中共筛选分离出表型突变家系145个。突变体类型分为4类,即氮营养缺陷型突变型、白化苗型、黄化苗型、植株矮小型。     

水稻抗白叶枯病T-DNA插入突变体侧翼序列的分离和分析

采用PCR-walking方法对水稻抗白叶枯病突变体G48的侧翼序列进行分析。通过特异性嵌套引物扩增和序列比对分析获得了该突变体T-DNA左、右边界侧翼序列,确定T-DNA插入水稻日本晴第三染色体clone OSJNBa0076E06(AC132215.1)位点上,左、右边界插入位点分别位于该克隆的第93 750、93 824碱基处。PCR-walking方法为T-DNA的侧翼序列分析提供了一种简单、高效的方法。     

水稻T-DNA插入氮营养缺陷型突变体的初步筛选

通过温室水培试验对根癌农杆菌介导的T-DNA随机插入构建的水稻突变体库的T1代2 173个家系进行筛选.以叶绿素SPAD值为指标,以苗期表观症状为参考指标,筛选到27个氮营养缺失的突变家系.对27个突变家系进行遗传分析、复筛及T2代鉴定,结果表明,编号955家系为氮营养缺陷型突变家系,且T2代能够稳定遗传.该材料通过进一步鉴定可用于分离、鉴定氮营养相关基因.     

甜瓜T-DNA插入突变体的构建

[目的]构建激活标签载体并将其转入甜瓜中,获得甜瓜T-DNA插入突变体,为甜瓜功能基因组学的研究奠定基础.[方法]用PCR法扩增目的DNA片段,经EcoRI和SacI顺序酶切,将目的片段连接到pCAMBIA2301载体上,构建含4 ×35s Enhancer DNA片段的激活标签载体.以甜瓜品种‘伽师’为材料,利用农杆菌介导的转化体系转化甜瓜愈伤组织,获得甜瓜T-DNA插入突变体.[结果]获得4株T0代甜瓜转化幼苗,通过Kan筛选和PCR鉴定甜瓜转化幼苗的DNA中是否含有目的基因,证明其中3株甜瓜转化幼苗为转基因植株.[结论]获得了突变植株,为甜瓜重要性状基因发掘奠定基础.     

T-DNA插入产生的水稻小粒突变体的遗传分析(英文)

[Objective] The aim of this study is to understand the genetic characteristics of a grain shape mutant and its possible role in genetic improvement of grain yield in rice. [Method] On the basis of the collection of T-DNA tag lines, the progeny of homozygous plants carrying T-DNA insertion were screened for mutants with mutated phenotypes. The genetic analysis of the mutant and test for the linkage between the mutated phenotype and the T-DNA insertion were carried out to determine its genetic characteristics. [Result] In the present study, a grain shape mutant induced by T-DNA insertion in rice was identified, which showed small grain. Genetic analysis of the mutant showed that the two types of phenotype, normal and small grain in the segregating populations derived from the T-DNA heterozygotes, fit the ratio of 3∶1. Test for Basta resistance showed that all the mutants were resistant while the normal plants segregated for resistant and susceptible by the ratio of 2∶1. The results indicated that the mutant phenotype cosegregated with Bar gene. The small grain mutant caused by T-DNA insertion was confirmed by PCR amplification aiming at T-DNA. [Conclusion] The grain shape mutant is useful for isolation of the tagged gene and genetic improvement in rice.     

一个水稻T-DNA插入类病斑突变体的初步研究

从T-DNA插入突变体中筛选到一个类病斑突变体AZT91,主要表现为生长缓慢、植株矮小、叶片出现条状褐斑,最后死亡。对突变体及其后代分离群体进行潮霉素抗性检测,证明该突变体是由T-DNA插入突变引起的,突变性状与T-DNA共分离。PCR和TAIL-PCR分析进一步证明了上述的观点。利用TAIL-PCR扩增了左边界侧翼序列,通过分析,初步推测该突变体可能是由于T-DNA插入后激活了单加氧酶基因的过量表达,破坏正常代谢途径,导致突变体死亡。该材料可用于水稻代谢调控机理的研究。     

插入T-DNA片段构建部分水稻突变体株系

 通过农杆菌Agrobacterium tumefaciens介导,将T-DNA片段高效插入水稻中花11的基因组DNA中。利用二次枝梗种子和未成熟种子的盾片愈伤组织作为转化起始物,探索并优化了影响愈伤组织诱导率、再生率和转化率的试验条件,得到近10 000株转化植株。经PCR分析和Southern 杂交证明已将外源基因整合到水稻基因组中。当代转基因植株可抗0.2%的除草剂Basta。200余棵T0代转化株系出现白化苗、宽叶、狭叶、黄叶、高秆、矮化并杂草化、不育、白化穗、皱穗和抽穗延迟等突变表型,结实率普遍较低     

水稻T-DNA插入氮营养缺陷型突变体的氮营养特征

利用营养液培养试验,研究水稻T-DNA插入氮营养缺陷型突变体的氮营养特性.结果表明,氮营养缺陷型突变体突变植株硝态氮和铵态氮吸收速率均低于原始亲本,植株氮含量及硝酸还原酶活性降低,氮同化能力下降,根系变短,根系体积及根系活跃面积变小,植株变矮.     

水稻Ds插入具芒突变体相关基因的功能预测

[目的]研究水稻Ds插入具芒突变体形成的分子机理。[方法]采用TAIL-PCR技术,从水稻Ds插入具芒突变体中克隆出Ds侧翼基因序列,分析被插入基因的结构,并分析预测基因编码的蛋白功能。[结果]Ds插入在具芒突变体7号染色体Os07g0588700基因前1 339 bp处。Ds插入位置的下游基因编码产物含一个锌指区CX2CX3FX5LX2HX3H,且含高度保守的QALGGH保守区,为水稻的单锌指蛋白。[结论]Ds转座元件插入基因组中,影响了编码锌指蛋白基因的表达调控,使突变体显示出具芒的表型。     

一个水稻生物产量突变体的遗传分析

突变体是研究基因功能的重要材料。除自然突变和物理、化学方法外,T—DNA插入诱变、转座子插入诱变等分子生物学手段也得到广泛的应用。而水稻全基因组测序的完成和水稻分子标记技术发展,使得通过图位克隆法从非插入突变的水稻突变体获得相关基因变得相对容易,目前,在水稻上通过图位克隆已克隆到一些重要的基因。该研究在转Bar基因水稻后代T1代群体中发展了一个高生物产量突变体,并对该突变体进行遗传分析,为该基因的克隆和功能分析提供参考。     

Genetic Analysis of a Biomass Mutant in Oryza sativa

[Objective] The study aimed to reveal the genetic model of a biomass mutant in Oryza sativa. [Method] In the process of screening and identification of Bar-transgenic rice,a biomass mutant was found in 10 lines of T1 progenies. The mutant was investigated for genetic analysis and agronomic traits by herbicide spraying and PCR amplification. [Result] The segregation ratio is consistent with mendelian law(3∶1). The mutant assumed not only higher plant height,wider straw and earlier florescence,but also more tillers,bigger spikes and resultantly higher biomass. PCR detections indicated that no co-segregation was observed between mutant traits and target gene(Bar) in the T-DNA inserted,proving that the mutant is not caused by the insertion of T-DNA containing target gene (Bar). [Conclusion] Our study may avail to understand the cloning of mutant gene and the mechanism of the mutant gene on biomass.     

Genetic Analysis of a Biomass Mutant in Oryza sativa

[Objective] The study aimed to reveal the genetic model of a biomass mutant in Oryza sativa. [Method] ln the process of seresning and iden-tification of Bar-transgenic rice, a biomass mutant was found in 10 lines of TI progenies. The mutant was investigated for genetic analysis and agronomic traits by herbicide spraying and PCR amplification. [Result] The segregation ratio is consistent with mendelian law(3:1). The mutant assumed not only higher plant height, wider straw and earlier florescence, but also more tillers, bigger spikes and resuhantly higher biomass. PCR detections indicated that no co-segregation was observed between mutant traits and target gene(Bar) in the T-DNA inserted, proving that the mutant is not caused by the insertion of T-DNA containing target gene (Bar). [Conclusion] Our study may avail to understand the cloning of mutant gene and the mechanism of the mutant gene on biomass.