花粉管通道法转化外源DNA的转基因技术

介绍了花粉管通道法的产生、分类及影响因素,阐述了花粉管通道法转基因技术的分子验证过程,列举了花粉管通道法在各作物育种上的应用,提出花粉管通道法应选择最佳导入方法与时期,从而提高外源DNA导人效率和遗传信息转化率,提高性状的变异频率,获得广泛的变异,提供丰富的变异材料。     

外源DNA花粉管通道途径导入机理研究进展

花粉管通道法由中国科学家提出,将基因工程和常规育种相结合,是一种颇具特色的转基因技术。详细地回顾了在花粉管通道技术转化机理研究方面的研究进展,并对花粉管通道技术的发展前景进行了展望。     

芝麻花粉在柱头上附着、萌发与伸长过程研究

以中芝14号为试验材料,采用荧光显微镜和石蜡切片法研究芝麻花粉在柱头上附着、萌发、伸长及在花柱内生长全过程。于授粉后不同时间段连续取样固定,并进行显微观察和数据统计,结果表明,在芝麻柱头上,授粉后5min基本无花粉萌发,10min有极少量花粉萌发,30min大量花粉开始萌发,并长出花粉管。花粉管具明显的绿色荧光,进入羽毛状柱头后继续生长于花柱基部至子房顶部的引导组织内。授粉后2h,生长最快的花粉管已经到达花柱基部,花粉管的平均生长速度约为3.0 mm/h。授粉后4h,大量花粉管进入子房。即使在授粉后24h,仍有部分花粉在萌发和生长中。     

芝麻花粉附着、萌发与花粉管伸长过程研究

以中芝14号为试验材料,采用荧光显微镜和石蜡切片法研究芝麻花粉在柱头上附着、萌发、伸长及在花柱内生长全过程.于授粉后不同时间段连续取样固定,并进行显微观察和数据统计.结果表明,在芝麻柱头上,授粉后5min基本无花粉萌发,10min有极少量花粉萌发,30min大量花粉开始萌发,并长出花粉管.花粉管具明显的绿色荧光,进入羽毛状柱头后继续生长于花柱基部至子房顶部的引导组织内.授粉后2h,生长最快的花粉管已经到达花柱基部,花粉管的平均生长速度约为3.0mm/h.授粉后4h,大量花粉管进入子房.即使在授粉后24h,仍有部分花粉在萌发和生长中.     

大豆育种中利用花粉管通道法导入外源基因应注意的几点问题

花粉管通道技术因其方法简便、易操作,目前在大豆遗传转化中普遍应用,笔者根据自己实际操作中得出的经验,提出了在大豆中利用花粉管通道法导入外源基因应注意的几点问题,并对该法的发展前景提出了几点展望。     

花粉管通道技术在棉花育种中的应用

利用花粉管通道法导入外源DNA技术是我国学者周光宇先生创建的 ,是我国基因工程研究中的一个特色。目前 ,花粉管通道法已在许多作物上得到了成功的应用。棉花花器大 ,繁殖系数高 ,尤其适用于采用花粉管通道法进行外源 DNA导入。目前我国培育和审定的转基因抗虫棉品种大多是以利用该技术获得的转基因抗虫材料为基础 ,通过系统选育或杂交选育而成的。山东棉花研究中心于 1 999年利用花粉管通道法将高产、抗病的海岛棉品系海71 2 4、异缘四倍体野生种达尔文氏棉及其他野生种系的叶片 DNA分别导入到陆地棉栽培品种石远32 1、鲁 735中 ,2 0 0…     

花粉管通道法将淀粉分支酶基因反义表达载体转入玉米自交系的研究

实验主要利用花粉管通道技术将淀粉分支酶基因的反义表达载体转入玉米自交系中,同时探讨了不同导入时间、不同DNA浓度、不同导入量等对其转化率的影响,并从中得出花粉管通道法的最佳转化条件。同时将转化的玉米植株进行PCR检测,初步证明外源目的基因已整合到玉米基因组中,并得到了转化植株的种子。     

早熟大豆外源DNA导入的RAPD分子验证

利用花粉管通道技术直接导入外源总ＤＮＡ，从而进行农作物品种改良，在国内外许多作物上已得到了广泛的应用。但外源总ＤＮＡ是否能够通过花粉管通道进入受体。后代的变异是不是由于外源总ＤＮＡ片段与受体基因组整合，表达所引起的，一直没有得到直接的分子生物学证据，本文利用ＲＡＰＤ这一分子生物学技术，对通过花粉管通导入外源总ＤＮＡ所获得的大豆早熟后代进行了分子验证。结果表明：在后代基因组中找到了供体具有而受体没有     

提高大豆花粉管通道技术的转化率研究

试验用荧光显微镜观察了大豆花粉的萌发。花粉管的延伸和进入子房的情况。从形态学和解剖学角度分析了大豆采用花粉管通道技术转基因成功率低的原因。并提出利用该技术进行转化的有利时间以提高转化率。     

花粉管通道法转化水稻研究进展

本文概述了花粉管通道法的转化原理，转化水稻了得的成果，外源DNA导入水稻后引起遥变异性状，转化水稻的操作方案，影响花粉管理法转化水稻的因素，用该法转化水稻的优点及存在的问题。     

芝麻细胞色素P450超家族的成员鉴定及SiCYP703A2的功能验证

细胞色素P450(CYP450)超家族广泛参与初生和次生代谢物质的生物合成,其中CYP703家族基因参与拟南芥、水稻等的花药发育.本研究利用生物信息学方法,从芝麻基因组中鉴定出303个CYP450基因,发现它们分为9个家族簇(clan),43个基因家族,随机分布于16个连锁群中.CYP703家族是71 Clan的成员之...     

芝麻种质营养品质分析及优质资源筛选

410份芝麻种质的分析结果表明,脂肪平均含量为53.13％,变异系数3.53％;蛋白质平均含量为26.39％,变异系数为7.03％。油酸、亚油酸含量总和占6种脂肪酸总量的80％以上。油酸和亚油酸对脂肪含量的直接效应分别为0.6907和0.9322。筛选出具有两种以上优质性状的品种19个,其脂肪、蛋白质、油酸、亚油酸含量分别超过56％、21％、42％、37％。     

简易超干燥芝麻种子贮藏方法的研究

采用木柜、干燥器和冰箱贮藏方法，对芝麻种子贮藏的效果进行了研究，结果以冰箱低温贮藏效果最佳，但成本高；其次是干燥器贮藏，干燥器内贮藏１７年的种子，含水量稳定在２．３％左右，粗脂肪、粗蛋白和脂肪酸组成含量未发生明显变化；田间种植出苗率仍在５０％以上，且能正常生长，无形态上的变异。     

黄淮海地区芝麻生产基地的确定

本文分析了影响黄淮海地区芝麻生产的主要生态因子,并采用聚类分析方法将黄淮海地区划分为芝麻最适宜种植区、适宜种植区、次适宜种植区和不适宜种植区,为该地区芝麻布局和商品基地建设提供依据。     

Sesame oleosin and prepro-2S albumin expressed as a fusion polypeptide in transgenic rice were split, processed and separately assembled into oil bodies and protein bodies

A recombinant polypeptide containing the precursor protein of a sesame storage protein, 2S albumin, fused to the C-terminus of a sesame oleosin was expressed in transgenic rice seeds under the control of a rice glutelin promoter. The recombinant polypeptide of 32 kDa, equivalent to the resultant molecular mass of sesame oleosin (15 kDa) and prepro-2S albumin (17 kDa), was detected in the endoplasmic reticulum fraction of maturing transgenic rice seeds, but not in the purified oil bodies or the soluble extract of transgenic seeds. However, sesame oleosin presumably fused with a 2 kDa C-terminal appendix originating from the signal sequence of prepro-2S albumin, was found in the purified oil bodies, and mature sesame 2S albumin apparently processed into two subunits (9 and 4 kDa) linked by disulfide bonds was detected in extracts of transgenic seeds. Immunogold labeling revealed that the sesame oleosin and 2S albumin were separately located in oil bodies and protein bodies of embryo cells of transgenic rice seeds. While sesame 2S albumin was also detected in protein bodies of endosperm cells of transgenic seeds, the co-expressed sesame oleosin, probably degraded due to the lack of oil bodies in this tissue, and was not detected. The results provide a new technique for introducing two recombinant polypeptides separately into rice oil bodies and protein bodies from one expression construct.     

Sesame oleosin and prepro-2S albumin expressed as a fusion polypeptide in transgenic rice were split,processed and separately assembled into oil bodies and protein bodies

A recombinant polypeptide containing the precursor protein of a sesame storage protein, 2S albumin, fused to the C-terminus of a sesame oleosin was expressed in transgenic rice seeds under the control of a rice glutelin promoter. The recombinant polypeptide of 32 kDa, equivalent to the resultant molecular mass of sesame oleosin (15 kDa) and prepro-2S albumin (17 kDa), was detected in the endoplasmic reticulum fraction of maturing transgenic rice seeds, but not in the purified oil bodies or the soluble extract of transgenic seeds. However, sesame oleosin presumably fused with a 2 kDa C-terminal appendix originating from the signal sequence of prepro-2S albumin, was found in the purified oil bodies, and mature sesame 2S albumin apparently processed into two subunits (9 and 4 kDa) linked by disulfide bonds was detected in extracts of transgenic seeds. Immunogold labeling revealed that the sesame oleosin and 2S albumin were separately located in oil bodies and protein bodies of embryo cells of transgenic rice seeds. While sesame 2S albumin was also detected in protein bodies of endosperm cells of transgenic seeds, the co-expressed sesame oleosin, probably degraded due to the lack of oil bodies in this tissue, and was not detected. The results provide a new technique for introducing two recombinant polypeptides separately into rice oil bodies and protein bodies from one expression construct.     

人工老化处理对芝麻种子生理生化特性的影响

以郑芝4号黑芝麻种子为材料,研究了人工加速老化处理对种子发芽指标﹑丙二醛(MDA)含量﹑过氧化物酶(POD)活性和超氧化物歧化酶(SOD)活性的影响。试验结果表明：随着种子老化加深,发芽势、发芽率、发芽指数﹑活力指数逐渐降低, 丙二醛含量逐渐升高。另外,老化种子内POD活性和SOD活性变化的总趋势是随老化时间延长而降低。由此认为芝麻种子人工老化及劣变的主要机制与抗氧化酶活性降低与和膜脂过氧化作用加剧有关。人工老化种子生活力出现快速下降的发芽势拐点是80%,其下降的趋势又与POD活性下降非常一致,POD可作为一个检测指标显示种子活力高低及丧失的特征。     

Devastation of sesame (Sesamum indicum L.) crops in different agroclimatic zones of India by genetically diverse subgroups of phytoplasma

The sesame crop is highly susceptible to infection by phytoplasmas, a class of cell wall-less plant pathogenic bacteria (Mollicutes), which is responsible for widespread loss of sesame crops in both North and South India in recent years. Therefore, characterizing the pathogen population is required before the control measures can be devised and implemented. With molecular tools based on nested polymerase chain reaction (PCR) assays, sequencing, restriction profiling, and phylogenetic analysis, phyllody-affected sesame plants collected from nine different states of India were found to be infected by phytoplasmas belonging to two 16Sr groups, namely 16SrI and II. Two subgroups of phytoplasma −16SrI-B and 16SrII-D— were prevalent in symptomatic sesame samples collected from North India, whereas phytoplasma of only the 16SrII group was found in South India. However, the latter samples were diverse, belonging to three different subgroups (16SrII-A, II-C, and II-D). In addition, yearly phyllody-affected sesame samples from Delhi for 4 consecutive years (2007–2010) showed variation in the infecting phytoplasma: the subgroup 16SrII-D was detected in samples collected in 2007, and 16SrI-B was predominantly found in the samples collected in the subsequent years. The study also provides molecular evidence for the association between 16SrI-B phytoplasma and different symptoms in sesame crops such as fasciation, little leaf, and stunting. This is the first study to report the association of the phytoplasma subgroups 16SrII-A and II-D with sesame crops in India. This study provides a baseline for designing specific detection and molecular analysis strategies for quarantine purposes. It also highlights the need for examining the dynamics of seasonal or location-specific variation in vector populations to determine the pattern of infection outbreaks.     

花生条纹病毒在芝麻上流行研究初报

调查和试验结果说明：感染ＰＳｔＶ的花生是芝麻黄花叶病毒病主要初浸染源．桃蚜传毒效率最高，为３５％，豆蚜、大豆蚜和棉蚜传毒效率均很低．病害流行与蚜虫发生密切相关．供试的１０个芝麻栽培品种均不抗病，但存在一定成株期抗性。自花生条纹病毒（ＰＳｔＶ）感染的芝麻病株上收获的种子，种植后未发现病毒种传现象。