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影响油菜游离小孢子培养再生因素的研究进展 总被引：1，自引：1，他引：1
游离小孢子培养已经成为油菜遗传育种和生物工程的重要内容。由于影响游离小孢子形态发生的因素较多，尤其是生长在大田环境下的植株，小孢子发育不一致，游离小孢子形态发生产生频率低，因此形态发生诱导率低仍然是限制该技术广泛应用的一个问题。为此，深入和全面了解影响小孢子形态发生能力及再生体系的各种因素，掌握这一领域的研究动态和今后发展的方向，有助于优化油菜小孢子的再生体系、促进油菜育种。本文总结了油菜游离小孢子形态发生的研究进展，并认为应用分子技术克服小孢子离体形态发生障碍有极大的发展潜力。 相似文献
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油菜小孢子培养技术体系研究 总被引：3，自引：0，他引：3
小孢子培养在油菜的基础研究和应用研究中均具有十分重要的意义。自1982年Lichter首次在甘蓝型油菜中进行小孢子培养获得成功以来,国内外在油菜小孢子培养技术方面已取得大量研究成果,包括油菜小孢子胚状体发生的影响因素,小孢子植株的再生、成苗、大田移栽、染色体加倍等,近年来又对一些关键技术环节加以了改进,笔者在对这些研究成果进行总结的基础上针对中国国情建立了大田条件下油菜高效小孢子培养技术体系。用该体系对甘蓝型油菜和新疆野生油菜的体细胞杂种后代进行小孢子培养的出胚率达到300枚/皿以上,采用小孢子苗直接移栽大田技术,成活率达到89.0%。此外还成功构建了含127个DH系的黄籽油菜DH群体及含115个DH系的粒重分离群体。 相似文献
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茄子组织培养与基因工程研究进展 总被引：8，自引：0，他引：8
本文综述了茄子组织培养与基因工程技术所取得的进展,并对茄子基因工程的发展进行了展望。迄今已利用茄子幼苗子叶、胚轴、叶片、胚、茎等外植体获得再生植株,是茄子离体再生研究最多的一个领域。其再生的关键取决于材料基因型、外植体类型、培养基类型等因素,不同基因型材料的再生能力差异很大,也影响再生的途径。原生质体是比较理想的进行遗传操作的受体,所以在其来源、培养液组成和成苗途径方面都有研究。花药和小孢子离体培养可以获得单倍体植株,节省获得纯系的时间,并有利于获得重组家系,为生产商用F1杂种开辟了一条重要途径。通过幼苗外植体器官再生、花粉(花药)培养、原生质体和小孢子培养已成功地建立了茄子受体再生系统。茄子基因工程技术在提高现有品质和创新种质方面发挥了积极的作用,农杆菌介化法是茄子转基因常用方法,已在外植体选择及再生、选择压筛选、农杆菌株系和转化浓度、预培养和共培养时间等影响因子方面做了大量的优化工作。转基因技术主要应用于茄子抗虫和单性结实基因工程方面并分别获得了一系列成果。在其他领域如抗病、抗逆等方面应用还很不足。因此作者认为未来的研究工作应集中于：目的基因的标记与克隆、高效受体再生系统的探索、更有效遗传转化方法的应用、扩大遗传转化范围,以提高实际应用水平。 相似文献
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雄核发育是小孢子或未成熟花粉细胞沿孢子体发育途径形成植株的过程，也是创制育种材料的有效途径之一。为建立重复性好、再生频率高的番茄小孢子诱导方法，文章通过回顾国内外研究进展，总结了番茄雄核番茄雄核发育各阶段（胚胎发生能力的获得、诱导小孢子分化形成胚状体以及胚状体再生形成单倍体植株）的主要影响因素，并简要归纳了现有的游离小孢子分离培养技术。分析表明，基因型是制约番茄小孢子诱导体系的关键因素，且需与发育时期、培养条件等多个影响因素同期发生，才能使雄核发育偏离配子体途径。最后，展望了番茄小孢子培养未来的研究方向与发展趋势，以期为番茄双单倍体技术的深入研究提供参考。 相似文献
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甘蓝型油菜小孢子培养中基因型及处理条件对成苗的影响 总被引：1，自引：0，他引：1
《种子》2020,(9)
为进一步提高甘蓝型油菜小孢子培养效率,研究了基因型对甘蓝型油菜小孢子胚产量和再生植株自然加倍率的影响,以及小孢子振荡培养过程中光照与后期低温处理对胚状体一次成苗率和再生植株自然加倍率的影响。结果表明,基因型对小孢子培养胚产量和再生植株自然加倍率影响较大,D 393的胚产量和自然加倍率很高,分别高达119.87胚·蕾~(-1)和79.87%,D 393的自然加倍率是D 524的2～3倍;光照对一次成苗率和再生植株自然加倍率没有影响,4℃低温处理15 d能提高小孢子一次成苗率0.8～3倍。本研究获得了一个胚产量和自然加倍率都很高的甘蓝型油菜DH系D 393;综合考虑一次成苗率和加倍效率,在甘蓝型油菜小孢子培养进程中振荡培养阶段可能不需要光照,而在转入B 5固体培养基前应该对胚状体进行适当低温和时长的处理,可提高小孢子培养效率。 相似文献
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《分子植物育种》2021,19(14):4745-4751
为进一步优化菜心游离小孢子胚再生技术体系,本研究以3份不同基因型的菜心小孢子胚状体为试验材料,研究不同培养基类型、培养基中不同琼脂浓度和不同胚龄对胚再生的影响,并对再生植株进行倍性鉴定。结果表明,使用B5培养基时各基因型的成苗率均高于MS培养基,褐化率低于MS培养基,B5培养基是菜心小孢子胚状体再生的适宜培养基;培养基中添加适宜浓度的琼脂可提高再生植株的成苗率,随着琼脂浓度的增加,出苗率呈先升后降的趋势,琼脂浓度为1.0%时,各基因型的出苗率达到最高;胚状体再生的最佳胚龄是25～29 d,胚龄时间过长或过短均不利于胚状体出苗,其中40 d以上胚龄的胚状体几乎不出苗;倍性鉴定发现菜心小孢子再生植株中同时存在单倍体、二倍体和多倍体,3个基因型的二倍体自然加倍率在62.22%～71.11%之间;75 mg/L秋水仙素处理可提高菜心再生株系的二倍体率。本研究为菜心游离小孢子胚再生体系优化及再生植株倍性鉴定提供了技术基础。 相似文献
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青麻叶大白菜小孢子培养及新品种选育 总被引：9，自引：1，他引：8
对影响青麻叶大白菜游离小孢子培养的因素进行了研究。结果表明：供体植株的基因型与小孢子胚胎发生密切相关;33℃,24h高温处理有助于小孢子转化成胚状体,每蕾成胚数比25℃恒温培养提高4．5倍;在培养基中加入外源激素和多种氨基酸可以提高胚状体的诱导效率,平均比对照增加59．64％;培养基的琼脂含量增加到12g／L能显著提高小孢子胚的成苗率,比含琼脂8g／L的高出40．5个百分点。通过对青麻叶大白菜小孢子胚的培养,获得二倍性的双单倍体植株,从中选出性状优良的纯系进行杂交组合的选配,培育出青麻叶大白菜的新品系。 相似文献
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为获得早熟型的双单倍体(DH)植株,加快早熟型红菜薹育种进程,本研究以15个早熟型红菜薹品种为试材,进行小孢子培养,观察小孢子发育过程并对再生植株进行倍性鉴定。结果表明,当花瓣/花药(P/A)长度比值为0.6～0.8时,红菜薹小孢子72%～78%处于单核靠边期;共有6个基因型红菜薹出胚,其中出胚率最大的为gy21-55,平均出胚率为14.3胚/蕾;选取子叶形胚进行植株再生,成苗率在90%以上;利用流式细胞仪对再生植株进行倍性鉴定,平均自然加倍率达76.8%。gy21-40、gy21-55表现出早熟特性,与对照植株相比获得的DH植株可分别提早5 d和8 d抽薹。 相似文献
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Haitham Sayed Hamed Kayyal Luke Ramsey Salvatore Ceccarelli Michael Baum 《Euphytica》2002,125(2):265-272
A total of 147 simple sequence repeat (SSR) markers (including86 barley and 61 wheat microsatellite markers) were tested for
their segregation in a doubled haploid (DH) and an F2 population of barley. The DH population consisted of 71 doubled haploid lines, developed from F1 plants of a cross between Tadmor and WI2291using isolated microspore culture technique. A genetic linkage map consisting
of 43 microsatellite markers was constructed using the DH population. Particularly on chromosome 4H microsatellite markers
showed distorted segregation ratios. Segregation of DH lines based on molecular markers were compared with segregation of
92 F2 lines from the same cross. The proportion of loci deviating from the expected monogenic segregation ratios in the DH population
was significantly higher (19/43loci, 44%) than in the F2 population (7/43 loci, 16%). The deviation was biased towards the WI2291 parent alleles. In line with this observation, WI2291
was found to perform better than Tadmor in regenerating green plantlets with the isolated microspore-culture technique.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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RAPD markers linked to a clubroot-resistance locus in Brassica rapa L. 总被引：10，自引：0，他引：10
Yasuhisa Kuginuki Hidetoshi Ajisaka Mamiko Yui Hiroaki Yoshikawa Ken-ichi Hida Masashi Hirai 《Euphytica》1997,98(3):149-154
Linkage of random amplified polymorphic DNA (RAPD) markers with resistance genes to clubroot (Plasmodiophora brassicae Wor.)
in Brassica rapa L. was studied in a doubled haploid (DH population obtained by microspore culture. Thirty-six DH lines were
obtained from F1 plants from a cross between susceptible ‘Homei P09’ and resistant ‘Siloga S2’ plants. ‘Homei P09’ was a DH line obtained
by microspore culture of the Chinese cabbage variety ‘Homei’, which is highly responsive in microspore culture. The resistant
line ‘Siloga S2’ was obtained by two rounds of selfing of the fodder turnip ‘Siloga’. Three RAPD markers, RA12-75A, WE22B
and WE49B, were found to be linked to a clubroot-resistance locus. These three markers were linked in the DH lines and an
F2 population and should be useful for marker-assisted selection in breeding programs.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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Alba Rivas-Sendra Manuel Campos-Vega Antonio Calabuig-Serna Jose M. Seguí-Simarro 《Euphytica》2017,213(4):89
We developed an eggplant doubled haploid (DH) population from a commercial hybrid through androgenesis in microspore culture. Morphological variation, reproductive ability and androgenic responsiveness were evaluated. The DH population showed segregation in vegetative traits related to leaf, flower and fruit, and in reproductive traits such as fruit and seed setting or germination rate. The DH population and subsequent generations also presented variation in the androgenic response, with null, low and high response lines. From this population, we were able to identify the first eggplant highly androgenic DH line (DH36), remarkably similar to the donor hybrid in terms of morphology and reproductive ability, but stably producing four times more calli than the hybrid. The segregating DH population is potentially useful for genetic studies and mapping of several traits, whereas the highly androgenic line DH36 may be used as a model line to facilitate the study of eggplant androgenesis and embryogenesis for both basic and applied research. 相似文献
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油菜小孢子培养影响因素及黄籽油菜双单倍体群体的构建 总被引：5，自引：0，他引：5
针对我国油菜小孢子培养供体材料一般种植在大田,受外界环境影响较大,出胚率较低的现状,对大田环境条件下小孢子培养的主要影响因素进行了研究。同时,为了构建黄籽油菜双单倍体群体,研究了黄籽油菜的出胚情况。结果表明,不同取样时间对小孢子胚状体再生频率的影响很大;秋水仙素处理游离小孢子对胚状体的诱导有一定的负面影响,浓度越高,影响越大;黄籽油菜的出胚率较低(0.56枚/皿),同样条件下,不到黑籽油菜的1/8。不同黄籽单株间出胚率差异较大,变异幅度在0.04~1.97枚/皿。采用小孢子苗直接移栽大田技术,移栽成活率达89.0%,经染色体加倍后,最终构建了含有127个株系的黄籽油菜双单倍体群体。 相似文献
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Efficient production of doubled haploid wheat plants by in vitro treatment of microspores with trifluralin or APM 总被引：4，自引：0，他引：4
Isolated microspore cultures from two doubled haploid (DH) lines of wheat, Triticum aestivum L., were used to develop an in vitro chromosome-doubling protocol. During the initial 24 h or 48 h of culture the microspores were treated with either of the two antimicrotubule herbicides trifluralin or amiprophos-methyl (APM) in concentrations ranging from 0.1 μM to 10μM. Untreated control cultures yielded 209 embryos per 100000 microspores, which is the equivalent of one spike. Among the regenerated plantlets 67% were green, and 15% of the flowering plants were spontaneously chromosome doubled. Treatments with both the herbicides had a significant effect on chromosome doubling, measured as the percentage of fertile regenerants. With the best combination of treatment duration (48 h) and herbicide concentration (10/μM) the percentage of fertile plants among regenerants could be increased up to 74% with APM and up to 65% with trifluralin. The largest numbers of DH plants per spike could be obtained with herbicide concentrations at 1–3 μM. Treatments with either herbicide at these concentrations resulted in an estimated average between the two genotypes of 27 DH plants per 100 000 microspores. These results demonstrate the high potential of APM and trifluralin as chromosome-doubling agents in isolated microspore cultures. The in vitro treatment integrated into tissue culture procedures will constitute an efficient method for chromosome doubling in future wheat breeding 相似文献
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Hong-hao Lv Qing-biao Wang Li-mei Yang Zhi-yuan Fang Yu-mei Liu Mu Zhuang Yang-yong Zhang Yu-hong Yang Bing-yan Xie Xiao-wu Wang 《Euphytica》2014,200(3):465-473
Cabbage Fusarium wilt (CFW), caused by Fusarium oxysporum f. sp. conglutinans, is one of the most devastating diseases of cabbage worldwide. In recent years, CFW has been spreading across northern Chinad. Thus, development of resistant cultivars is necessary to control CFW. Here we report microspore culture from a cross of two elite cabbage inbred lines, 96–100 (highly resistant to CFW) and 01–20 (highly susceptible to CFW), in order to obtain doubled haploids (DHs) to cultivate cabbage materials for resistance breeding. A total of 196 DHs were obtained in 2011 and 2012 and three elite DH lines were selected from them through agronomic trait evaluation as well as marker-assisted selection. These three elite DH lines were crossed with elite cabbage inbred lines to create hybrids, which were then evaluated for their resistance and agronomic traits. Three excellent hybrids were selected. The use of microspore culture and marker-assisted selection greatly shortened the time required to obtain these excellent hybrids. These resistant varieties may become an effective control of CFW in diseased areas. 相似文献
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Information from previously published studies that are basic to this study is: ) Following isolated barley microspore culture around % of the resulting barley plants are completely fertile genetically homozygous doubled haploids 《分子植物育种》2007,5(2):280-281
Information from previously published studies that are basic to this study is： 1） Following isolated barley microspore culture, around 80% of the resulting barley plants are completely fertile and genetically homozygous doubled haploids （DH） （Kasha et al., 2003）. 2） Chromosome doubling occurred during early stages of microspore culture, mainly by nuclear fusion （Kasha et al., 2001）. 3） A fairly synchronous population of microspores was selected to determine the cell cycle stages （G1, S, G2） by relative DNA density measurements （Shim and Kasha, 2003a）. 4） The pretreatments of spikes used to induce microspore embryogenesis influenced the cell cycle progression during pretreatment. （i） Using a combination of cold （4℃） and 0.3 mol/L mannitol for 4 days, the microspores were held at the G1 uninucleate stage. （ii） The often used pretreatment of cold （4℃ for 21 to 28 d） permitted slow microspore stage progression so that most cells were at the G2 stage or had become binucleate by the end of the pretreatment. （iii） Pretreatment with mannitol for 4 days at 25 ℃ did not block cell cycle progression but prevented cell wall formation after the 1^st mitotic division, leading to nuclear fusion and chromosome doubling. 相似文献