赤霉素、光照及基因型对花生体细胞胚诱导和植株再生的影响

【目的】完善花生离体再生繁殖体系,为花生的遗传转化提供技术支持。【方法】以桂花系列花生品种为材料,以胚小叶为外植体,MS＋10mg/L2,4-D为体细胞胚诱导基本培养基,MS＋3mg/L6-BA＋0.8mg/LNAA＋（0～15mg/L）GA3为苗诱导基本培养基,研究光照、赤霉素和基因型对花生体细胞胚诱导和植株再生的影响。【结果】在光暗比14∶10条件下,花生体细胞胚诱导率明显高于全黑暗培养,体细胞胚诱导率增加7.5～37.5个百分点。在不同花生品种中,桂花26体细胞胚诱导率最高,达62.8%,桂花833最低,为21.7%。在成苗培养基中添加5～15mg/LGA3利于提高体细胞胚的成苗率。桂花26和桂花771在5mg/LGA3处理下成苗率最高,分别为42.8%和35.3%;桂花836、桂花1026、桂花833在15mg/LGA3作用下成苗率最高,分别为38.7%、33.3%、26.4%。所有品种与GA3组合中,以桂花26与5mg/LGA3组合成苗率最高,其次是桂花836与15mg/LGA3组合。【结论】适宜的光照条件有利于花生体细胞胚的发生;成苗培养基中添加GA3利于促进体细胞胚诱导成苗;桂花26和桂花836是体细胞胚诱导植株再生较理想的材料。     

Field performance of embryogenic cell suspension-derived banana plants (Musa AAA, cv. Grande naine)

Growth and yield characteristics of two different clones of banana plants (Musa AAA cv. Grande naine) originating from four months old embryogenic cell suspensions were studied. These characteristics were compared with those plants produced by the conventional in vitro budding multiplication method. Two types of variants were observed during the acclimatization phase among 500 embryogenic cell suspension derived plants. The first type related to banana plants with `variegated or deformed leaves' were also observed in in vitro budding derived plants. The second type concerned `fasciated-leafed' plants. During the field growth, these two variant types produced plants morphologically similar to the other plants. Thus, none of the cell suspension derived plants exhibited off-type traits in the field. A Fisher block model was used to compare the field performances of the two clones produced through the two in vitro propagation techniques. The analysis of variance showed that there were no significant differences between the plants produced by either micropropagation techniques for the plant height and circumference, the length of the reference leaf, the number of nodal clusters of the inflorescence and of fruits, the bunch weight, the period of time between planting and flowering, and between planting and harvesting. This study showed that banana plants with an agronomical behaviour similar to those produced by the conventional in vitro budding method could be regenerated from embryogenic cell suspension. This revised version was published online in July 2006 with corrections to the Cover Date.     

Efficient shoot regeneration and somatic embryogenesis from immature cotyledons of Brassica napus L.

Immature cotyledons of Brassica napus cv.‘Topas’ were used to investigate the effects of various naphthaleneacetic acid and benzylaminopurine concentrations on morphogenesis in vitro. A high efficiency of adventitious shoot regeneration and somatic embryogenesis was achieved on Murashige and Skoog medium supplemented with the above growth regulators. Two types of somatic embryogenesis were obtained, the first directly and the second via a callus stage.     

Relationship between somaclonal variation and type of culture in cucumber

Highly inbred B line of cucumber was used to compare the effect of four types of in vitro culture on somaclonal variation. The plants were regenerated from the following types of culture: twelve- and eighteen-month-old liquid culture of meristematic clumps (LMC₁₂₍₁₈₎), ten-month-old embryogenic cytokinin-dependent suspension (CDS), eighteen-month-old embryogenic cytokinin-dependent suspension in medium with modified NH⁺ ₄/NO₃ ^- ratio (CDS 1.7), twelve-month-old embryogenic auxin-dependent suspension (ADS), thirty six-month-old embryogenic auxin-dependent suspension in medium with modified NH⁺ ₄/NO₃ ^- ratio (ADS 1.7) and recurrent leaf callus regeneration (RLC) – repeated 5 times. The differences in the incidence of the following properties were observed: the ploidy of R₀ plants, the segregation of new morphological traits in R₁ and the germination ability of R₁ seeds. R₁ families with the segregation of new phenotypes were most numerous in CDS (62.5%) and LMC₁₈ (57.9%), next in CDS1.7 (35.7%), while the smallest number was found in LMC₁₂ (11.1%) and RLC (3.4%).Tetraploid and mixoploid plants occurred in ADS1.7 and ADS (100%) whereas CDS and RLC were observed to contain only tetraploids, respectively 33.3% and 55.2%. There were no changes of ploidy after LMC₁₂, LMC₁₈ and CDS1.7. Among new phenotypes there were such that have not been described so far in cucumber: ginkgolike leaf (gll), yellow-green chlorophyll mutants (y-gc), serrate margin of corolla in male and female flowers (smc). This revised version was published online in August 2006 with corrections to the Cover Date.     

A comparison of the somaclonal variation level of Rosa hybrida L. cv Meirutral plants regenerated from callus or direct induction from different vegetative and embryonic tissues

Summary Flowering plants of Rosa hybrida L. cv Meirutral have been obtained either from direct regeneration of adventitious shoots on leaf and root fragments, or through organogenesis and somatic embryogenesis on calli derived from anther, ovule, petal, sepal, receptacle, leaf, stem internode, root and zygotic embryo tissues. The calli derived from floral parts exhibited rhizogenesis. In this case direct induction of adventitious shoots from selected roots had to be performed in order to generate plants. A histological study of the morphogenetic calli was carried out. The plants regenerated directly and those regenerated from calli of leaf, stem internode, root and zygotic embryo tissues, together with reference plants propagated by cuttings, were compared on a phenotypic basis by taking into account petal number, form and colour, and plant growth habit. From these observations, it can be concluded that directly regenerated plants are as stable as reference plants while plants regenerated from callus are unstable, especially those derived from zygotic embryo tissues.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-Dichlorophenoxyacetic acid - GA3 gibberellic acid - IAA 3-indole-acetic acid - MS Murashige and Skoog - NAA 1-naphthalene acetic acid     

Somatic embryogenesis from floral tissue of cassava (Manihot esculenta Crantz)

The aim of this study was to examine the embryogenic potential of floral material of the cassava cultivar MCOL 1505. Macerated immature inflorescences were found to be highly embryogenic, with almost 78% of the explants producing somatic embryos. Somatic embryos were also produced from whole male florets and half florets although at much lower rates. No regeneration was obtained from anther, microspore or floret wall tissue. Somatic embryos derived from immature inflorescences were regenerated via organogenesis and the plants derived from this process were assessed in terms of phenotype and ploidy level. If haploid plants could be produced by this method, this would have significant implications in assisting traditional cassava breeding, as this would allow homozygosity to be reached more rapidly. In a crop such as cassava, which is highly heterozygous in nature, the use of haploids in a breeding programme could considerably shorten the time taken to produce new desirable cultivars. This is the first report on plant regeneration through somatic embryogenesis from floral tissue of cassava. This revised version was published online in July 2006 with corrections to the Cover Date.     

TDZ对橡胶树花药愈伤组织诱导和体细胞胚发生的影响

为了提高橡胶树优良品种的愈伤组织诱导和体细胞胚发生的频率,促进其花药植株再生体系的优化,以橡胶树品种‘云研73-477’、‘热垦525’和‘云研77-2’的花药为外植体,研究不同浓度的TDZ对愈伤组织诱导和细胞胚胎发生能力的影响。结果表明,在愈伤组织诱导实验中,MS+TDZ 0.2 mg/L+ 6-BA 1 mg/L+2,4-D 1.5 mg/L+NAA 0.5 mg/L+椰子水50 mL/L+蔗糖70 g/L效果最好,诱导率均达到最大,分别为98%、98.67%、28.6%,当TDZ浓度逐步升高时,其诱导率又有所下降。在体细胞胚诱导实验中,愈伤组织诱导阶段加入TDZ对体细胞胚的发生有抑制作用;将愈伤组织转移到不含TDZ的培养基中继代增殖后,体细胞胚的诱导率随继代次数的增加呈上升趋势;愈伤诱导阶段使用的TDZ浓度越低越利于体细胞胚的分化;在体胚诱导阶段以TDZ代替KT后,体细胞胚诱导率较低,且分化出的体胚为畸形胚。这表明TDZ虽然有利于愈伤组织的产生和生长,但不利于体细胞胚的发生。     

生物技术在柑橘砧木上的应用

本文概述了当前国内外柑橘砧木生物技术尤其是基因工程技术和体细胞杂交技术在柑橘砧木种质改良和品种选育的研究现状与进展,可为我国柑橘砧木资源的研究与开发利用提供参考。基因工程在柑橘育种中的应用起步较晚,但近年来发展迅速,通过运用农杆菌介导法、附体腋芽转化－离体扩繁鉴定法、DNA直接导入法等基因工程技术对培育新型抗性柑橘砧木品种取得了一定的成果;在体细胞杂种的运用方面,通过原生质体融合,获得了大量具优良性状的杂种体细胞,并建立了多个抗病杂交后代群。目前,柑橘砧木生物技术的研究方兴未艾,现代生物技术在柑橘砧木资源多样性研究与种质改良方面仍是今后的主攻方向。     

棉花gDNA体细胞染色体FISH技术

介绍了棉花基因组ＤＮＡ（ｇｅｎｏｍｅＤＮＡ,简为ｇＤＮＡ）体细胞染色体荧光原位杂交〔ＦＩＳＨ〕的技术流程,并着重分析和讨论了影响试验结果的关键因素,包括染色体和探针的变性条件、染色体的蛋白酶Ｋ处理技巧等。试验中作为靶ＤＮＡ的体细胞染色体采用棉属异源四倍体种海岛棉;探针和封阻均采用ｇＤＮＡ,材料是棉属二倍体种Ａ染色体组（Ａｇｅｎｏｍｅ）的棉种（亚洲棉和草棉）和Ｄ染色体组（Ｄｇｅｎｏｍｅ）的棉种（瑟伯氏棉、雷蒙德氏棉、戴维逊氏棉等）,分别交互使用。试验结果比较理想,获得良好的ＦＩＳＨ片子,而且重复性好。