小叶杨原生质体培养植株再生及其同工酶的变化

从小叶杨（Ｐｏｐｕｌｕｓｓｉｍｏｎｉｉ）胚性悬浮细胞分离得到大量有活力的原生质体，产量高达３．８×１０７ｇ－１ＦＷ。高密度（３×１０６／ｍＬ）液体浅层培养可促进小叶杨原生质体的分裂和生长；以葡萄糖为唯一碳源的Ｋｍ８ｐ培养基，有利于原生质体的持续分裂；在培养基中添加有机氮，有助于提高原生质体的植板率。原生质体形成细胞团后，逐步降渗和分皿培养，能有效促进细胞团的持续分裂。愈伤组织在不同分化培养基上的分化频率存在着明显的差异，同工酶分析说明，分化能力强的愈伤组织，其ＴＣＡ循环和磷酸戊糖途径较为活跃，ＧＯＴ、ＥＳＴ和ＰＥＲＯＸ的活性比较高。当芽伸长到２—３ｃｍ时，从基部切下转移到无激素的１／２ＭＳ培养基上诱导生根并形成完整植株。移入盒内，在温室生长良好。     

花药特异嵌合启动子的构建及雄性不育转基因拟南芥的获得

为了克服TA29／Barnase转基因雄性不育植物的温度敏感性，在TA29启动子上游串联了CaMV35 S启动子，同时在CaMV35 S启动子上游串联一个调节序列antherbox，构建成嵌合启动子antherbox-CaMV35 S-pTA29。该启动元件调控的GUS基因瞬间表达实验表明它是花药颈毡层特异的启动子。这个启动元件及其调控下的barnase基因构建植物表达载体转化，拟南芥菜，获得了雄性不育的转基因植株。     

诱导根瘤菌与稻类根系结瘤共生

1986年水稻原生质体再生植株获得成功以来,国内外对原生质体再生植株及其后代的性状变异已有一些报道,1988年中国科学院遗传研究所培育籼稻品种大谷早原生质体再生植株成功,1988-1990年我们对大谷早再生植株及其后代R_26个株系360植株的形态性状变异,进行了系统的田间     

紫花苜蓿原生质体培养与植株再生

由和田苜蓿和单选苜蓿叶肉分离原生质体，采用改良的K8P和MS培养基进行液体浅层培养。原生质体经一次分裂，二次分裂和多次分裂，形成小细胞团，并很快长成小愈伤组织。愈伤组织在MS分化培养基上增殖并分化，产生小植株，经MS无激素培养基生根形成完整的再生植株。     

雄性不育基因工程及其在蔬菜育种中的应用

本文概述了雄性不育系在杂交制种中的重要地位和雄性不育基因工程的主要策略 ,讨论了近年来我国利用雄性不育基因工程创制蔬菜雄性不育系的进展 ,并探讨了这一技术在蔬菜育种上的应用前景     

利用RAPD标记对彩色棉遗传多样性的分析

以新彩棉1号、2号和71份彩色棉材料为基础,利用RAPD标记对彩棉品种之间分子标记遗传差异进行研究。从240个随机引物中筛选出10个引物能在73份彩棉材料间扩增出稳定性较好的多态性片段。73份彩棉材料的欧氏遗传距离在1.00～4.79之间。按类平均距离法进行聚类分析表明,这些彩色棉材料可分为7组,品种间的遗传关系与品种自身的系谱有关,大多数品种的遗传基础比较狭窄。     

小麦花粉管途径转化及高效筛选体系的建立

本文报道了一种从大量花粉管途径转化处理后代中快速有效地筛选到转基因植株的方法。转化质粒为含bar筛选标记基因的天麻抗真菌蛋白(GAFP)与兔防御素(NP-1)双价抗病基因植物表达载体(pBI35S-gafp-NP1-bar),受体小麦品种为扬麦158和Alondra。对花粉管途径转化处理获得的种子分3步进行筛选鉴定：首先,转化处理种子密播于塑料盘中,待小苗长到3叶期时,用浓度为120mg／L的Basta除草剂弥雾喷施筛选,约10d后绝大部分小苗变黄枯死,只有约1％的高抗Basta除草剂的T0小苗存活;然后对其Basta抗性植株用gafp基因引物进行PCR检测,其中62．5％的Basta抗性植株为PCR阳性;最后将T0代PCR阳性植株后代种子(T1)分别发芽并按株系混合取样提取DNA,分别用gafp和bar基因引物进行PCR检测验证,结果均为阳性。表明外源基因已整合入小麦基因组并由T0代植株稳定遗传到T1代,同时证明该筛选方法是可靠有效的。     

观赏植物色香形墓因工程研究进展──文献综述

综述了花卉色、香、形三大重要性状的基因工程修饰的最新研究进展。着重介绍了花色基因工程修饰的策略、方法以及培育人类梦寐以求的蓝花月季等的可能性。     

西瓜子叶原生质体再生愈伤组织的获得

从西瓜(Citrullus vulgaric)无菌苗子叶游离原生质体,用改良的DPD培养基和琼脂糖珠包埋方法培养,可以得到再生细胞的高频率分裂,继而得到原生质体再生愈伤组织。     

大豆微卫星诱发突变的分子分析

Microsatellite or SSR marker is an efficient tool for plant genotype identification, molecular mapping and marker-assisted selection. Objective of this study is to analyze the mutagenized microsatellite variations in soybean genome and reveal nature of these mutations. In the present study, mutations at fifteen microsatellite loci were detected in genomic DNAs of soybean mutant E182 induced by EMS (ethyne metyl sulfate) using PCR amplification of 485 pairs of SSR primers. These fifteen mutagenized microsatellite loci with repeat number variation were Satt005, Sattll7, Satt185, Satt282, Satt290, Satt420, Satt452, Satt483, Satt569, Satt579, Satt600, Satt602, Sat-086, Sat-107 and Sat-135, respectively. Sequencing results of these fifteen loci indicated that microsatellite sequences at Satt282, Satt483, Satt579, Satt600 and Satt602 loci were respectively deleted 1 -, 3 -, 8-, 20 - and 1 - trinucleotide (all [ATT]1-20 except for [CAA]8 [TAA]12 at Satt600 locus) repeats, which made allele sizes at these five loci decrease 3, 9, 24, 60 and 3 bp, respectively. And while microsatellite sequences at the other ten mutated loci, Satt005, Sattll7, Satt185, Satt290, Satt420, Satt452, Satt569 and Sat-086, Sat-107, Sat-135, were respectively inserted 1-, 6-, 6-, 3-, 4-, 3-, 8- trinu-cleotide repeats (ATT)1-8 and 12-, 6-, 16- dinucleotide repeats (AT)6-16, making allele sizes at these ten loci increase 3, 18, 18, 9, 12, 9, 24, 24, 12, 32 bp, respectively. On the other hand, eleven events of base mutations were detected in flanking regions at seven (Sat- 107, Satt185, Satt282, Satt420, Satt569, Satt579 and Satt600) of fifteen mutated microsatellite loci. These base mutations consisted of 6 transitions (4T→C and 2 A→G), 2 transvertions (A→T and T→A), 1 insertion (T) and 2 deletions (A and T). The experimental results proved that EMS mutagenesis could cause different types of mutations at microsatellite multilocus in soybean genome, including repeat number variations in microsatellite regions and random base mutations in flanking regions. We found three mutational biases, which were frequent insertion mutations of repeat units, initiating positions of microsatellite sequences of repeat unit insertions/deletions and both flanking-base T ↓ A of these insertion/deletion positions. In addition, the resolution capacity of high-quality agarose gels was sufficient to distinguish differences of only three base pairs in this experiment.