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油菜下胚轴高频率植株再生因素研究 总被引:34,自引:0,他引:34
有关油菜的组织培养已有许多报道,但多局限于采用不同外植体获得再生植株,对于影响外植体高频率再生的诸因素未作系统分析。在油菜的基因转化中外植体的来源和再生方式都直接影响着转化效果。根据Mukhopadlyay(1992)等人和Radke(1992)的工作,下胚轴是较好的外植体来源,而直接成苗的再生方式最大限度地保存了原种质的遗传特性。我们对芥菜型油菜下胚轴高频率植株再生的因素进行了研究,以期为油菜基因转化工作中高效外植体再生系统的建立提供依据。 相似文献
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杜仲叶片为外植体的植株再生 总被引:2,自引:0,他引:2
杜仲不仅是传统中药植物,也是重要的胶源植物,具有很高的经济价值。本研究采用生长21天的杜仲无菌苗叶片为材料,基本培养基为MB(MS无机盐,B5有机物)。不定芽的分化:切取杜仲叶片(叶盘)为外植体,在附加6-BA(1.0mg/L),IAA(0.5mg/L),KT(2.0mg/L),TDZ(0.2mg/L)的MB培养基上培养一个月,分化得到不定芽,分化率高达到56%。壮芽和芽的增殖:不定芽培养在MB 6-BA(1.0mg/L) NAA(0.05mg/L)的培养基上,可使不定芽迅速得到增殖。生根和移栽:将切取的单芽培养在1/2MS IBA(1.5mg/L) 蔗糖20g/L的(培养基上,一个月后,从芽的基部生长出根,获得杜仲再生植株。本研究通过叶盘法建立杜仲植株再生体系,为杜仲的遗传改良奠定了基础。 相似文献
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高频率生菜植株再生及转化体系的建立 总被引:8,自引:0,他引:8
通过对两种生菜及其不同外植体在培养基上不定芽发生能力的筛选,得到“大湖366”叶切片不定芽发生率高达87%的培养体系,并且丛生状不定芽可陆续发生。将叶切片与农杆菌共培养,对经km筛选后所得再生株进行X-Glu染色,兰色反应阳性率为50%。 相似文献
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龙牙百合的植株再生与遗传转化 总被引:19,自引:0,他引:19
以龙牙百合鳞片叶切块为外植体,通过多种培养基的比较试验,得出MS 2,4-D 2mg/L 6-BA 0.2mg/L是诱导愈伤的最佳培养基,MS 6-BA 1mg/L NAA 0.05mg/L是不定芽诱导及伸长的适宜培养基。MS N_AA2mg/L是生根的最佳培养基。本试验已建立了适用于龙牙百合遗传转化的快速高频再生系统。用携带有几丁质酶基因和β—1、3葡聚糖酶基因的工程菌,通过农杆菌介导法和基因枪转化法转化龙牙百合,经PCR和点杂交检测证明外源基因已经整合到植物染色体中。同时对农杆菌介导法和基因枪法进行比较,发现农杆菌介导法的转化率为16.7%,基因枪法的转化率为50%,因此可能基因枪转化法更适于龙牙百合的遗传转化。 相似文献
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【目的】探究影响农杆菌介导棉花体细胞转化体系产生的再生植株育性的影响因素。【方法】从载体大小、目的基因大小、不同阶段组织培养时间等因素对6个载体的转基因再生植株当代(T0)育性影响进行分析。【结果】不同载体/目的基因的转基因植株不育率呈现显著差异,但是转基因植株育性与载体大小、目的基因大小之间没有明显关系。进一步研究发现转基因再生植株不育率与胚性愈伤组织分化-植株再生的培养时间呈显著正相关,分化-植株再生培养时间少于110 d、110~130 d、130~150 d、150~170 d、超过170 d的植株不育率分别为19.6%、45.5%、63.8%、72.3%、94.5%;而与诱导愈伤组织形成-胚性愈伤组织分化的持续培养时间没有相关性。【结论】转基因棉花再生植株育性受到胚性愈伤组织持续培养时间的显著影响,缩短胚性愈伤组织到植株再生的培养时间能够提高转基因再生植株的育性。 相似文献
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影响长寿花离体培养及植株再生的几个因素 总被引:5,自引:2,他引:5
对长寿花茎段培养7~8d,部分茎段腋芽萌动。30d培养结果表明,芽诱导的最佳培养基是:MS+6-BA1.0mg?l-1+NAA0.5 mg?l-1;芽伸长的最佳培养基是:1/4MS;形成有效节的最佳培养基是:MS。继代培养中各培养基上无叶茎段均较带叶茎段萌芽率高。在5、6、7号培养基上,叶片培养9d时,从叶柄基部开始形成不定芽,在其它培养基上,14d才开始有不定芽形成。30d培养结果还表明,诱导叶片再生不定芽的最佳培养基是:1/2MS+6-BA1.0 mg?l-1。诱导根生长的最佳培养基是:1/4MS。 相似文献
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Evidence of functional unreduced gametes in Trifolium repens L. 总被引:4,自引:0,他引:4
The current research reports the first evidence of functional 2n gametes in megaspores of Trifolium repens L. Reciprocal backcrossing
of a colchicine doubled hexaploid (2n = 6x = 48) T. repens × T. nigrescens (6x H-6909-5) F1 hybrid to T. repens produced four seeds from 1700 pollinations. The backcross involving 6x H-6909-5 as the female parent
and T. repens as the male resulted in only one seed which was, as expected, a pentaploid (2n = 5x = 40) presumably carrying
four genomes of T. repens and one genome of T. nigrescens. On the other hand, those backcrosses which utilised T. repens as
the female parent and 6x H-6909-5 as the male parent, resulted in 3 viable seeds. These were unexpectedly all heptaploid (2n
= 7x = 56) with presumably six genomes of T. repens and one genome of T. nigrescens. The occurrence of heptaploids from a
T. repens × 6x H-6909-5 (4x–6x) cross can only be explained by the union of n (= 3x = 24) pollen from 6x H-6909-5 with 2n
(= 4x = 32) eggs from T. repens.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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Protoplasts were isolated from leaves of Lotus corniculatus and Trifolium repens and from cotyledons of L. corniculatus. These protoplasts divided and produced colonies. The plating efficiency of protoplasts of both species was improved when agarose was used as a supporting medium. Plants were regenerated more regularly and in larger numbers from colonies of L. carniculatus than T. repens. The use of a culture line of T. repens that had been selected for its response in culture markedly increased the proportion of protoplast-derived cultures which regenerated shoots. One regenerant of T. repens (P6) was analysed for morphological and cytalogical variation. This plant was abnormal and highly aneuploid with a wide range of chromosome numbers. 相似文献
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为了探讨白三叶(Trifolium repens L.)种质资源农艺性状变异和相关性,揭示各种质材料的特征特性和种质资源群间的遗传关系,本研究对78份白三叶种质的12个主要数量性状的变异和相关性进行了分析。结果表明:这些种质材料存在着丰富的遗传多样性,各个数量性状间均呈正相关,植株高度与小叶长度、小叶宽度、小叶大小、叶柄长度呈显著正相关;小叶长度与小叶宽度、小叶大小呈极显著正相关,与叶柄长度、叶柄直径呈显著正相关;小叶宽度与小叶大小呈极显著正相关,与叶柄长度、花柄直径呈显著正相关;小叶大小与叶柄长度、叶柄直径、花柄直径呈显著正相关。该批种质材料能为各类白三叶育种和遗传研究提供较为丰富的亲本材料。 相似文献
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为探索低温胁迫对白三叶的叶绿素、可溶性糖和丙二醛等生理指标的影响,为白三叶的抗寒性育种和资源的合理利用提供基础数据,试验以新品系08-2和海法白三叶(Trifolium repens L.‘haifa’)为材料,依次设置10℃、5℃、0℃、-5℃、-10℃、-15℃等6个处理,研究低温下叶绿素、可溶性糖和丙二醛等的变化,对材料的抗寒性进行评价。结果表明,变温对白三叶的叶绿素、可溶性糖和丙二醛等的影响较大。随着温度的降低,白三叶的叶绿素含量总体呈现降低的趋势,在较低(≤0℃)温度处理下,08-2的叶绿素含量降低速度小于海法的;08-2的可溶性糖含量在0℃时达到最高,海法的在5℃达到最高;丙二醛含量和电导率逐渐升高,但08-2变化幅度低于海法。综合分析表明,新品系08-2耐寒性优于海法白三叶。 相似文献
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Nitrogen Fixation by Hybrids of White Clover (Trifolium repens L.) and Trifolium nigrescens 总被引:1,自引:0,他引:1
M. T. Abberton J. H. MacDuff A. H. Marshall T. P. T. Michaelson-Yeates 《Journal of Agronomy and Crop Science》1999,183(1):27-33
Development of hybrids between white clover ( Trifolium repens L.) and Trifolium nigrescens provides a novel route for genetically improving the reproductive capacity of white clover, provided the hybrids are agronomically viable, particularly with respect to N2 fixation. A comparative study of growth and rates of N2 fixation over 21 days was conducted with the parental species, F 1 hybrids and backcross hybrids, in flowing solution culture, without a supply of mineral N to the plants. T. nigrescens was unable to fix N2 in association with the strains of Rhizobium leguminosarum biovar. trifolii selected for inoculation. Rates of N2 fixation per plant increased in the order T. nigrescens < F 1 hybrid < T. repens < backcross 1. Specific rates of N2 fixation (days 0–21) increased in the order T. nigrescens < F 1 hybrid < backcross 1 < T. repens . Dry matter production and nodule biomass per plant increased at a higher rate in backcross 1 hybrids than in T. repens. The results suggest that the potential for N2 fixation by backcross 1 hybrids is at least as great as that by T. repens . 相似文献
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M. T. Abberton T. P. T. Michaelson-Yeates A. H. Marshall K. Holdbrook-Smith I. Rhodes 《Plant Breeding》1998,117(5):494-496
Hybrids between the stoloniferous Trifolium repens (2n = 4x = 32) and the rhizomatous Trifolium ambiguum (2n = 4x = 32) have been produced using T. repens as the recurrent parent. Morphological characteristics of the parent species and the F1, BC1, BC2 and BC3 hybrids were examined in a glasshouse experiment. Leaflet ratios and general plant shape indicated that the BC3 hybrid was similar to T. repens. Separation of plants into components of above- and below-ground growth showed that T. repens had a greater total plant dry weight than T. ambiguum but 24% of its dry weight was in roots compared with 22% in roots and 44% in rhizomes in T. ambiguum. The BC3 generation contained plants that were predominantly T. repens-like with stolons, but also had a small proportion of their total dry weight (3%) as rhizome, confirming the potential to produce plants that combine stoloniferous and rhizomatous growth. The proportions of rhizome and stolon varied within the BC3 generation, enabling farther selection and plant breeding. The BC3 hybrids produced similar numbers of inflorescences to T. repens however, fertility was lower, although there was some variation within the BC3 for both characters. 相似文献
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Inbred lines of white clover (Trifolium repens L.) have been produced utilising the rare self-fertility (Sf) allele. Twenty-two
lines of four distinct groups have been maintained through five generations of selfing by single seed descent. Fourteen lines
were used in crosses to produce F1s. Both parents and hybrids were analysed for a range of morphological characters. Positive heterosis for dry matter production
was observed in half the hybrids. No other trait showed significant heterosis. The degree of heterosis appears to be related
to the extent of variation in morphological characters between the parental lines, some combinations of lines yielding heterotic
hybrids in all cases, others in none. The majority of the F1 hybrids are superior in terms of dry matter production to eight control varieties.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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Introgression as a Route to Improved Drought Tolerance in White Clover (Trifolium repens L.) 总被引:1,自引:0,他引:1
A. H. Marshall C. Rascle M. T. Abberton T. P. T. Michaelson-Yeates & I. Rhodes 《Journal of Agronomy and Crop Science》2001,187(1):11-18
Backcross hybrids between the important forage legume white clover Trifolium repens , which is stoloniferous, and the related rhizomatous species T. ambiguum have been produced using T. repens as the recurrent parent. The drought tolerance of parents and two generations of backcross plants, possessing both stolons and rhizomes, was studied in deep soil bins in a glasshouse both as monocultures and in mixtures with perennial ryegrass. Soil moisture content, leaf relative water content, and leaf water potential were measured on plants subjected to drought cycles of 4 weeks duration and those watered normally and maintained at field capacity. Six cycles of drought, carried out over 2 years, showed that T. ambiguum and the first and second generation backcross hybrids maintained a greater leaf relative water content and higher leaf water potential than T. repens at comparable levels of soil moisture. The dry matter yield of T. repens was significantly reduced over each drought cycle compared to the watered treatments, while the yield of T. ambiguum increased over the 4-week cycle in both treatments. The yield of the backcross generations was intermediate between those of the two parental species. The implications of these results for white clover germplasm improvement programmes are discussed. 相似文献
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Nitrogen Fixation in Hybrids of White Clover (Trifolium repens L.) and Caucasian Clover (Trifolium ambiguum M. Bieb) 总被引:2,自引:0,他引:2
M. T. Abberton J. H. MacDuff S. Vagg A. H. Marshall T. P. T. Michaelson-Yeates 《Journal of Agronomy and Crop Science》2000,185(4):241-247
Hybrids between white clover (Trifolium repens L.) and Caucasian clover (or Kura clover, Trifolium ambiguum M. Bieb) are a potential route for the improvement of drought tolerance and persistence in white clover. However, to be agronomically viable they must show no significant reduction in their potential for nitrogen fixation relative to white clover. A comparative study of growth rate and nitrogen fixation was carried out in flowing solution culture without a supply of mineral nitrogen to the plants. The two parental species and two generations of backcross hybrids, with white clover as the recurrent parent, were assessed. The growth rate and N content of T. ambiguum were significantly lower than those of the other lines. However, dry matter production, nodule biomass per plant and rates of fixation were similar in second‐generation backcross plants and white clover. The results suggest that the agronomic potential of this novel germplasm is not compromised by limitations with respect to nitrogen fixation. 相似文献