花粉管通道法转化大豆的分子表征

采用花粉管通道技术将含有gus基因的pBI121质粒DNA导入辽豆14，从处理的100朵花中获得53粒种子。经PCR检测53粒种子的幼苗，共检测出6株阳性植株。采用RT—PCR和GUS组织化学染色法对所获得阳性植株进行检测，其中2株幼苗(T0代17号和33号)均出现阳性结果，进一步对这2株阳性植株进行Southern blot检测，同样获得了阳性结果。上述检测结果表明，gus基因已整合到染色体上并得到了表达。对转基因植株后代进行了PCR跟踪检测，结果从17号株系的28株中检测出19株阳性植株，33号株系的35株幼苗中检测出17株阳性植株，表明gus基因可以在转基因植株后代遗传。本研究结果为大豆花粉管通道转化技术提供了比较充分的分子生物学证据。     

花粉管通道法转化蓖麻的研究

为验证花粉管通道法进行蓖麻转基因研究的可行性,优化花粉管通道法导入方式及导入时间,为花粉管通道法转基因技术在蓖麻分子育种的应用提供参考依据。采用荧光显微观察对蓖麻品种通蓖5号人工授粉后花粉萌发及花粉管生长情况进行了研究,结果表明,花粉管通道法转化蓖麻的最佳时间为授粉后6～24 h。利用花粉管通道技术,将含有GUS基因的植物双元表达载体pPZP221 SGN导入蓖麻品种通蓖5号。对85株T0植株进行PCR检测,得到10株阳性转化植株,阳性率约为11．8％。     

花粉管通道法在玉米转基因育种中的应用

花粉管通道法是上世纪70年代提出并用于改造植物材料的基因转化方法,目前在很多农作物上有所应用。本文对该法在玉米自交系改良上的研究内容、成果进行综述,并对其今后在玉米品种改良方面的进一步应用做了展望。     

花粉管通道法转化小麦影响因素的研究

以西农1376和中13小麦品种(系)为受体,携带叶片衰老抑制基因PSAG12-IPT的pCMLA35-1质粒为转化载体,对花粉管通道转化法中的基因型、转化时间、质粒DNA 浓度进行了研究。结果表明：不同基因型、转化时间对结实率影响不显著,质粒DNA浓度对结实率影响显著。在0~700 ng/μl范围内,小麦结实率随着质粒DNA浓度的增加而降低,其中质粒DNA浓度300 ng/μl和500 ng/μl与对照相比结实率差异达到显著水平,而700 ng/μl则达到极显著差异。不同基因型的转化率因转化时间和质粒DNA浓度的不同而有所差异,其中西农1376的适宜转化时间为40~80min,适宜质粒DNA浓度为300~500ng/μl,以500ng/μl×40min处理的转化率最高,而中13小麦的适宜转化时间为80~120min,适宜质粒DNA 浓度为300ng/μl。     

Bar基因的玉米花粉管通道法转化

通过花粉管通道法利用抗除草剂基因（bar）转化3个玉米自交系,D0代获得35株GUS阳性植株．其中17株PCR阳性植株,除草剂筛选获得抗性植株13株。研究发现DNA溶液浓度影响转化率,当浓度为100μg／ml时,转化率最高,为2．61％。DNA导入时间距人工授粉时间的延长,结实率升高,但转化率显著下降,人工授粉后1d导入DNA产生的转化株最多,为19株,平均转化率为1．35％。     

外源DNA导入花粉管通道技术的发展和应用

本文综述了花粉管通道法的创立与发展,以及应用此方法所取得的一系列研究成果,对遗传转化中所涉及的机理和分子验证进行了探讨,并且分析了影响花粉管通道法导入外源基因的因素,进一步比较了该方法较其它常用的转化法的优势与局限性。     

花粉管通道法转Bar基因胡麻后代的分子检测

以花粉管通道法转Bar基因胡麻T1代胡麻为研究对象,用CTAB法提取DNA,经PCR分别扩增外源基因Bar基因的片段,扩增产物用琼脂糖凝胶电泳进行检测分析,筛选Bar基因成功整合到基因组的单株,结果在114个T1系亚麻植株中检测出5株含有Bar基因的阳性亚麻植株,转化率为4.4%。探讨了花粉管通道法转Bar基因在亚麻中的应用前景。     

利用花粉管通道转化谷子DNAj基因获得转基因小麦

为提高小麦的抗旱能力,采用花粉管通道法将本实验室克隆的谷子抗逆相关基因DNAj转入小麦中.对授粉方式、柱头处理方式、质粒DNA浓度及DNA稀释试剂等因素对结实率的影响进行了对比,结果只有授粉方式对结实率的影响有显著差异,其他条件下结实率均未显示显著差异.对转化获得的703株T0小麦植株提取DNA进行分子检测,有1株小麦稳定扩增到了预期的1 260 bp的目的基因片段.虽然转化效率偏低(1.42‰),但说明利用花粉管通道法将谷子的抗逆相关基因DNAj导人小麦是可行的,为创造抗逆性改良的小麦新材料奠定了基础.     

采用花粉管通道法将蛋白激酶基因导入玉米自交系的研究

通过花粉管通道法将ZmPtil,ZmPtil-1,ZmCIPK2三种蛋白激酶基因植物表达载体分别导入玉米自交系吉444,经草丁膦筛选后得40株抗性植株,通过PCR检测其中28株为PCR阳性植株,平均转化率为1.33％,最后收获11株转基因植株.干旱条件下通过对转基因T1植株的单株籽粒产量、千粒重两个指标进行差异显著性和...     

花粉管通道转基因技术及在水稻分子育种中的应用

中国学者创立花粉管通道法转基因技术以来,在多种农作物分子育种中得到应用。广泛的研究表明,花粉管通道法转基因技术与其他转基因方法相比,具有能直接得到转化种子,转化频率高,操作简便经济,大量快捷,性状稳定快和导入材料多样性等优点。同时重点介绍了花粉管通道法转基因技术在水稻分子育种中的研究和应用,采用该方法将多种植物外源DNA导入水稻,获得了包括植株形态,产量性状,抗病虫性,稻米品质和生理生化等方面的变异,是一种简便有效的水稻分子育种方法。     

Bar基因的亚麻花粉管通道法转化

[研究目的]为鉴定T1代转基因亚麻植株中抗除草剂基因(bar)整合是否成功。[方法]通过花粉管通道法将bar基因导入转化植株,以叶片涂抹法对T1代转化植株进行田间草丁膦最低致死浓度筛选;以最低致死浓度草丁膦进行田间喷施后筛选耐受草丁膦药液的抗性植株,并用PCR检测bar基因阳性的抗性植株数目。[结果]草丁膦对T1代转化植株的最低致死浓度为100 mg/L。得到耐受草丁膦药液的抗性植株44株,PCR检测结果表明,8株为PCR阳性植株,平均转化率为18.2%。[结论]花粉管通道法将bar基因成功转入亚麻基因组。     

Assignment of the denso Dwarfing Gene to the Long Arm of Chromosome 3(3H) of Barley by Use of RFLP Markers

RFLP analysis of 105 doubled haploid lines from a cross between the barley varieties ‘Magnum’ and ‘Goldmarker’ located the denso dwarfing gene on the long arm of chromosome 3(3H), approximately 8 cM distal to the RFLP locus Xpsrl70. Lines with the denso gene showed a distinctive prostrate juvenile growth habit and tended to have later ear emergence times and lower plant grain weights, ear grain weights and 50 grain weights.     

Some aspects of cross-pollination in wheat (Triticum aestivum L.) 1. Pollen concentration in the field as influenced by variety,diurnal pattern,weather conditions and level as compared to the height of the pollen donor

Summary For the large-scale production of hybrid seed the cross-fertilizing capacity of wheat must be increased. Breeding good pollinators is, therefore, a necessity. Five spring wheat varieties were examined for differences in pollen shedding capacity under normal field conditions. The examination was carried out by collecting pollen on object slides every hour from 06.00 till 20.00 h during flowering. In general, the variety Peko proved to be the best shedder of pollen, while Opal was the worst. Gaby, Jufy I and Orca were intermediate. Consistent results were hard to obtain mainly due to varying weather conditions.It was established that pollen is released throughout the whole period of daylight. The maximum release of pollen, however, occurred from 08.00 till 11.00 h. A less pronounced and much smaller peak was noticed from about 15.00 till 18.00 h. Most pollen appeared to be released at a temperature of 16–20°C and a relative humidity of 70–75%. Although there seems to be some relationship between the diurnal pattern of the pollen release and the indicated values of these meteorological factors, one cannot say that the former is entirely determined by the latter.It has been suggested in the literature that a male-sterile parent that is shorter than its pollinator would be most likely to produce good seed set in the hybrid wheat seed production field. Investigations showed that a greater concentration of pollen was found below the ear level of pollinator plants than at or above ear level.     

Resistance and susceptibility to tobacco mosaic virus (TMV) in the tomato,Lycopersicon esculentum,demonstrated by histological differences in the development of stem tissue

Summary Seedlings of two tomato varieties, one susceptible and the other resistant to TMV, were inoculated at the first true leaf stage with a mixed inoculum of TMV and PVX. PVX was added to induce an active and uniform reaction. The seedlings were then grown in flats in the greenhouse, and plants were harvested each day following the initial inoculation for a period of eighteen days. The stems of these plants were fixed, embedded in paraffin, sectioned and stained. Examination of the different cross sections revealed distortions in the development of the vascular system of the susceptible variety.