马铃薯叶肉原生质体再生植株的研究

马铃薯两个品系小叶子x多子白和乌盟601的叶肉原生质体在原生质体培养基中诱导出愈伤组织,叶肉原生质体来源的愈伤组织转移到MS＋2mg/1ZT+0.1mg/1 IAA培养基中,培养至70天以后,开始发生芽的分化,待芽生长到2－3cm高度时,转入MS＋0．05mg/1 NAA培养基中,很快出根长成完整植株,带1－2片叶的茎段移栽入灭菌的混合土壤中生长并结出薯块。     

马铃薯野生种叶肉原生质体培养及其植株再生（简报）

马铃薯野生种叶肉原生质体培养及其植株再生（简报）何亚文,李耿光,张兰英（中国科学院华南植物研究所,广州５１０６５０）ＭＥＳＯＰＨＹＬＬＰＲＯＴＯＰＬＡＳＴＣＵＬＴＵＲＥＡＮＤＰＬＡＮＴＬＥＴＲＥＧＥＮＥＲＡＴＩＯＮＦＲＯＭＷＩＬＤＳＰＥＣＩＥＳＯＦＰ...     

细叶黄芪叶肉原生质体植株再生

从细叶黄芪(Astragalus tenuis)外植体愈伤组织分化出的再生苗叶片分离原生质体。原生质体培养在改良 K8p 培养基中形成了愈伤组织。增殖后的愈伤组织转入分化培养基中分化出苗。幼苗在生根培养基中长出不定根,再生成为完整植株。再生苗叶肉原生质体在 AY培养基中,种子无菌苗叶肉原生质体在改良 K8p 或 AY 培养基中均不能形成愈伤组织。较低的2,4-D 浓度有利于原生质体愈伤组织的形成和分化,过高的2,4-D 浓度对愈伤组织的形成和分化有不利的影响。     

可乐茄叶肉原生质体培养再生植株

本文报道茄属果树可乐茄（ＳｏｌａｎｕｍｑｕｉｔｏｅｎｓｅＬａｍ．）叶肉原生质体的分离、培养及植株再生。幼嫩叶片原生质体经酶游离、纯化后,以１×１０４个／ｍｌ密度培养于稍加改良Ｋ８ｐ（附加２,４－Ｄ０．５ｍｇＬ（－１）、ＮＡＡ１．０ｍｇＬ（－１）和ＢＡ０．５ｍｇＬ（－１））的培养基中,三天后开始分裂,一周分裂３—４次。一个月形成小细胞团,植板率为０．１—０．２％,小细胞团转培养于ＭＳ＋２,４－Ｄ０．５ｍｇＬ（－１）上增殖后进行分化。原生质体来源愈伤组织在ＩＡＡ（０．１—１．０ｍｇＬ（－１））与ＢＡ或ＺＴ组合的培养基中能诱导器官发生,芽分化率最高可达４２．９％;但ＩＡＡ、ＢＡ、ＺＴ三者一起使用未见任何器官分化。小芽在ＭＳ＋ＩＡＡ０．２ｍｇＬ（－１）中生根成植株。可乐茄叶肉原生质体的植株再生,可应用于育种和茄属植物遗传工程研究。     

叶肉细胞原生质体培养再生植株变异的研究

为了满足社会对经济作物在数量及质量上不断提高的要求,寻找克服远缘杂交不亲和性及定向改变植物遗传性状的新途径,以培育出适合要求的优良品种,一直是生物学工作者努力的一个重要方面。近来年的研究结果表明,熟练地掌握植物原生质体培养的技术和方法,对于利用原生质...     

草木樨状黄芪叶肉原生质体的植株再生

分别从草木樨状黄芪胚轴再生苗的上部和下部叶片分离原生质体。来自上部叶片的原生质体培养在P_2培养基(含2,4-D 1.0mg/L)中获得了较高的分裂频率(48.9%)和愈伤组织再生频率(321块/m1),过高和过低的2,4-D对于愈伤组织的再生都是不利的。来自下部叶片的原生质体分裂频率很低,不能形成愈伤组织。小愈伤组织转入固体或液体增殖培养基中均能快速生长。愈伤组织转入分化培养基或继续在液体培养基中振荡培养均能分化出芽,频率达100%。目前已获得了大量的再生植株,部分已移栽成活。     

茼蒿叶肉原生质体培养再生植株

从茼蒿(Chrysanthemum coronarium)第1片真叶制备原生质体,经悬滴培养和琼脂糖小块培养形成愈伤组织。愈伤组织在分化培养基上诱导芽的分化,再经诱导生根得到再生植株。     

桑树叶肉原生质体培养再生植株

近年来,木本植物原生质作诱导再生植株的研究越来越受到国内外学者关注。但在林木树种中,迄今成功的种类仍然不多,在文     

百脉根愈伤组织原生质体再生植株

百脉根无菌苗幼茎在含２．０ｍｇ／Ｌ－,２,４－Ｄ,０．１ｍｇ／Ｌ２－ｉｐ的ＭＳ培养基上诱导和继代培养愈伤组织。选取绿色松散颗粒愈伤组织分离原生质体。原生质体培养在调整珠ＫＭ８Ｐ,Ｖ－ＫＭ,ＭＳ和ＳＨ培养基上「含３００ｍｇ／Ｌ,ＣＨ,２％ＣＷ,２％蔗糖,６％葡萄糖,２．０ｍｇ／Ｌ,２,４－Ｄ,０．５ｍｇｇ／Ｌ,ＢＡ,５ｍｍｏｌ／Ｌ ＭＥＳ」,原生质体再生细胞均能分裂,并形成小愈伤组织,但以ＫＭ８０为     

Studies on the effect of protoplast density and genotype mixing on cell regeneration

Genotypes of Solanum tuberosum, assessed for their differential response to a protoplast regeneration protocol, were identified. This enabled comparisons to be made in relation to their abilities to regenerate into dividing cells and colonies. Protoplasts from genotypes with contrasting regeneration responses were plated as single or mixed genotype cultures at various plating densities in replicated, randomised experiments, and the effect on the regeneration of particular cell types observed. Protoplasts from single genotype cultures showed intergenotypic variation in the extent of cell regeneration and there were significant effects of genotype mixing and density × genotype mixing interactions. Overall, the effect of mixing was beneficial to a regenerating culture but significant density genotype mixing interactions showed only a positive benefit at the lowest plating density. The protoplast mixing phenomenon was not correlated with the behaviour of the same genotypes at the plantlet level in experiments with in vitro meristem cultures.     

利用一步基因中断法构建克鲁氏乳酸酵母leu2突变体

将克鲁氏乳酸酵母ＬＥＵ2　基因编码区的部分序列用酿酒酵母的ＵＲＡ３基因替换,然后用此段转化两株克鲁氏乳酸酵母。通过体内同源重组,部分缺失的外源leu2　片段取代下了酵母染色体上的正常的ＬＥＵ2基因,由此得到leu2转化子。经过这些转化子在非选择条件下的稳定性测定,没有发现回复子。结果表明,用一步基因中断法成功地建立了稳定的ＬＥＵ2基因突变体。这为在克鲁氏乳酸酵母中构建有效的宿主-载体系统提供了一个有用的营养缺陷型选择标记。Acstract: TheＬＥＵ2 gene of the yeast Kluyveromyces lactis was disrupted by replacing a part of the coding sequence with URA3 gene of the yeast S.cerevisiae. Transformation of two k.lactis strains with the disrupted leu2 fragment resulted in the substitution og partilly deleted LEU2 gene for the with-type LEU2 gene on the chromosome.Thus, two leu2 mutants were generated and no reversion could be detected after prolonged growth in the non-selective medium. The results show that the stable leu2 mutants have been constructed successfully by one-step gene disruption. The isolation of these mutants would provide a useful auxotrophic marker to facilitate the development of an efficient host-vector system in K.lactis.     

Plant regeneration from mesophyll protoplasts of Solanum commersonii dun

Somatic fusion of Solanum commersonii, a frost tolerant wild potato species not crossable with Solanum tuberosum, relies on the possibility to isolate and culture protoplasts. This study was conducted to determine whether protoplasts could be isolated and plants regenerated in three S. commersonii accessions. Shoot cultures for protoplast isolation were maintained on Murashige and Skoog medium. Mesophyll protoplasts were isolated and cultured using a protocol originally described for S. tuberosum with some modifications. Differences were evident among the three accessions for protoplast yield, plating efficiency and regeneration frequency. Protoplast yield ranged from 3.0 to 8.5 × 10⁶ protoplasts per g of fresh tissue. At 1–2 × 10⁴ protoplasts ml⁻¹, which was the optimal plating density, 10–20% of plated protoplasts gave multicellular colonies. Regeneration of shoots was observed in two accessions only, the maximum regeneration frequency being 66%. In one of these accessions the reduction of sucrose concentration in regeneration media improved the regeneration frequency from 14 to 35%. About three hundred plants were rooted in vitro and successfully transferred to soil.     

基因组对芸苔属作物原生质体培养及植株再生的影响

本文以包心菜、芜菁油菜、浙油６０１的无菌苗叶肉原生质体为材料,经不同液体培养基浅层培养,细胞分裂并形成愈伤组织。愈伤组织经增殖后,转到分化培养基上诱导分化,均获得了再生植株。本文着重研究了植物基因组对原生质体分裂频率及植株再生的影响。研究结果表明：（１）植物基因组对原生质体分裂频率的影响随原生质体培养基的不同而异;（２）植物基因组对原生质体再生植株影响显著,芜菁油菜的Ａ基因组不利于原生质体再生植株     

Changes in abscisic and gibberellic acids contents during the release of potato seed dormancy

Gas chromatographic measurements demonstrated that the content of endogenous gibberellic acid increased and that of abscisic acid decreased during storage of potato seeds, suggesting that the dormancy of the seeds is controlled by the balance between these two hormones. This revised version was published online in July 2006 with corrections to the Cover Date.