水稻愈伤组织再分化培养方法的优化

在水稻愈伤组织再分化培养基中添加不同浓度配比的6-BA、iP、NAA三种植物生长调节物质,组成34种不同的幼苗分化培养基,对水稻愈伤组织进行再分化培养。结果表明：（1）培养基中添加2mg／L 6-BA＋2mg／L iP＋0.2mg／LNAA有利于水稻愈伤组织形成绿点;（2）采用三步分化法,对于已分化出绿点的水稻愈伤组织,能明显提高成苗率。     

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。     

大蕉幼雄花易碎胚性愈伤组织长期继代培养及其植株再生

大蕉未成熟雄花接种到胚性愈伤组织诱导培养基中,4～5个月后可诱导出胚性愈伤组织,并可在继代培养基上长期增殖。这些继代培养了6年多的松软易碎的胚性愈伤组织转移到含有0.2 mg/L 6-BA的分化培养基中,可诱导出芽,得到了53株再生植株,这些再生植株可进一步扩大繁殖。组织学切片证明长期继代培养的愈伤组织维持了胚性的状态。     

Efficient In Vitro Shoot-regeneration Systems in Cassava (Manihot esculenta Crantz)

Two different protocols for in vitro regeneration of cassava using zygotic embryos and nodal axillary meristems have been developed. In both cases, buds were regenerated directly from excised explants without an intervening callus phase after a two-step culture procedure. In cotyledonary explants derived from zygotic embryos, prolific shoot formation occurred within 2—3 weeks on MS medium supplemented with 0.5—5 mg/1 BAP alone or in combination with 0.1 mg/1 NAA. Nodal explants with axillary meristems derived from aseptically grown seedlings or stem cuttings were used to initiate a round compact bulb-like structure on MS medium containing 10 mg/1 BAP. These latter structures, when cultured on MS medium supplemented with 0.1 mg/1 NAA, 1 mg/1 BAP and 0.1 mg/1 GA₃, produced multiple shoots. Somatic embryos isolated at the globular/torpedo stage from zygotic embryo explants were also capable of multiple shoot production on medium with 1 mg/1 BAP. Rooting of regenerated shoots exceeded 95 % in phytohormone-free MS medium. No change in their ploidy levels was observed. Therefore, the protocols developed should be of use in the particle gun and Agrobacterium-mediated genetic transformation of cassava.     

陆地棉体细胞胚的白化与植株再生的关系

通过对棉花体细胞胚胎发育过程的观察发现，其发育的四个时期即球形期，心形期，鱼雷期和子叶期均有白化现象。成熟期的白色叶胚比绿色子叶胚在成苗培养基上生长快，且形成再生植株的频率高。     

Potential,constraints and applications of in vitro methods in improving grain legumes

Grain legumes, the important constituents of sustainability‐based cropping systems and energy‐limited vegetarian diets have long been the subject of scientific research. Tremendous technological strides were made in the so‐called orphan crops, in terms of both varietal improvement and generation of basic information. Despite recalcitrancy and high genotype dependency, in vitro culture techniques such as organogenesis, in vitro mutagenesis, embryo rescue and in vitro gene transfer have been deployed for improvement of several grain legumes and these played an important role in introgression of desirable genes from related and distant species and creation of additional genetic variability. Stable and reproducible regeneration protocols resulted in the development of genetically modified chickpea, pigeon pea, cowpea, mungbean, etc., while embryo rescue was deployed successfully for recovery of interspecific recombinants, a few of them exploited for the development of commercial cultivars. Nevertheless, doubled haploidy witnessed limited success and protoplast regeneration and in vitro mutagenesis remained of academic interest. The present review focuses on the progress, achievements, constraints and perspectives of using in vitro technology in grain legume improvement.     

聚乙二醇细胞融合技术在牛手工体细胞克隆中的应用初探

本研究分3个实验。实验1对比了不同浓度的离子霉素对牛去透明带卵母细胞孤雌激活的效果;实验2比较了手工半卵切割法去核与显微半卵切割法去核的效果;实验3对聚乙二醇（PEG）细胞融合技术在牛手工体细胞克隆的应用进行了初步探讨。结果表明,对于去透明带牛卵母细胞孤雌激活,离子霉素浓度为2．0μmol／L组的激活效果最好,其囊胚发育率（29．4％）极显著地高于浓度为5．0μmol／L的对照组和0．5μmol／L组（囊胚率分别为8．3％和9．4％,P〈0．01）;采用半卵切割法去核,手工切割的半卵存活率（85．6％）与显微操作（90．3％）差异不大,切割速率（约100枚卵／h）相仿。对双半卵与体细胞进行同步融合处理时,应用50％浓度的PEG融合率（39．3％）极显著好于45％、55％和60％的PEG（融合率分别为0％、16．2％和5．3％,P〈0．01）。     

新疆野生枸杞愈伤组织分化及植株再生

以新疆枸杞的叶片、茎尖基部为外植体,分别诱导愈伤组织、继代培养和诱导分化,结果表明:①不同外植体愈伤分化率有明显差异:叶片愈伤组织分化率最高为100%,茎尖基部愈伤组织分化率最高为80%。每块愈伤的芽分化数最多可达15～20个。②激素组合影响愈伤分化。诱导分化最适培养基为6 BA2.0mg·L-1 IAA0.5mg·L-1,KT0.5mg·L-1 NAA1.0mg·L-1。③将分化出的无根苗转接到MS无激素培养基上,生根率可达到85%以上。因此叶片为诱导愈伤组织分化的最适外植体。     

10个特选水稻品种的离体培养特性

将10个特选的水稻品种的离体培养特性与模式品种台北309进行了比较．试验发现在培养基含有2mg/L的2,4-D的情况下添加0．5mg/L的BA对爪哇稻胚性愈伤组织的形成显著有利,而对其他品种的正向作用不大或反向作用明显．虽然华航1号和华航3号的愈伤组织和台北309一样能够在没有细胞分裂素的培养基上再生植株,但再生效率很低．在含有BA的培养基上所有品种的愈伤组织都能够再生出更多的植株,添加铜元素对部分品种的植株再生也有促进作用．先将愈伤组织进行分化预备培养然后才进行植株再生培养能够取得最好的植株再生培养效果．本研究结果表明,采取适当的方法能够显著改善大多数品种的培养反应．经过比较筛选出离体培养特性与模式品种相似的优质籼稻品种水晶占和台湾香占,可以作为利用生物技术进行品种改良的材料．     

蒲桃胚胎发育及多胚来源的初步研究

观察了蒲桃花果的形态结构和胚胎发生.蒲桃胚囊的发育属蓼型.每个蒲桃子房内有10多个胚珠,但成熟果实内多数只有1粒种子,少数为2粒,原因为一些胚囊不能发育成熟,而那些成熟的胚囊又有一部分不能顺利地完成双受精.蒲桃胚乳发育属核型.在受精之前,多数珠心的表皮下即有珠心胚原细胞出现,这些细胞分裂成为不定胚的原始体.随着某些珠心内的胚囊败育,这些不定胚原始体也归于消亡.而那些顺利完成双受精的胚囊外的不定胚原始体则可往胚囊方向靠拢,并进一步侵入胚囊.通常,侵入胚囊的珠心胚达5—9个.此外,也观察到了裂生多胚现象.最终,每粒发育成熟的种子内存有2-10个胚,合子胚、裂生胚和珠心胚混杂在一起.