白杄体细胞胚悬浮培养的动力学研究

白(ＰｉｃｅａｍｅｙｅｒｉＲｅｈｄ.ｅｔＷｉｌｓ.)是我国特有的云杉属树种,在林业生产和环境绿化中均具有重要地位。其体细胞胚胎发生的研究,一方面可用于优良种质的大规模快速繁殖,为植树造林和园林绿化提供优质苗木;另一方面可作为遗传转化的再生系统,进行树种遗传…     

柳树遗传学研究现状与前景

柳树种类繁多,分布广泛,是重要的阔叶树种,其遗传学研究也较早受到重视。该文从柳树性状遗传变异、群体遗传、遗传鉴定、遗传图谱构建和QTL定位几个角度介绍了柳树遗传学的研究进展,并结合木本植物的特点和柳树的生物学特性进行分析,认为柳树是理想的林木遗传学研究材料,可作为模式树种,应受到特别关注。     

柳树遗传学研究现状与前景

柳树种类繁多, 分布广泛, 是重要的阔叶树种, 其遗传学研究也较早受到重视。该文从柳树性状遗传变异、群体遗传、遗传鉴定、遗传图谱构建和QTL定位几个角度介绍了柳树遗传学的研究进展, 并结合木本植物的特点和柳树的生物学特性进行分析, 认为柳树是理想的林木遗传学研究材料, 可作为模式树种, 应受到特别关注。     

林木群体遗传多样性和多位点遗传结构

本文评述了群体同工酶基因位点遗传结构研究中的主要度量方法及其在森林树种中的应用概况。群体遗传结构可由单位点和多位点两类参数度量。 文中对这两类度量及其关系分别进行了讨论。阐明了多位点遗传结构的信息对林木改良和资源保存策略的重要意义。     

杨树重要品种( 无性系) 的AFLP 指纹分析

品种的准确鉴定及其遗传相关性的了解对杨树育种和品种管理具有非常重要的意义。本试验采用AFLP对来自青杨组和黑杨组的21 个重要杨树品种( 无性系) 的鉴定与遗传相关性进行了研究。结果显示, 筛选的4对AFLP 引物总共产生了181 条多态性带,尤其是每对引物对每个品种都产生了独特的指纹图谱; 聚类分析和多维尺度分析将试验材料大体上分为五类, 结果不仅显示了组间不同品种的差异, 而且大体上区分了我国原生品种和外来品种。本研究表明, 所有品种都可被筛选的引物准确鉴定, 遗传相关性的推断结果与它们的系谱或分类基本一致。另外, 本研究还表明AFLP 技术完全可用于大规模地构建杨树树种DNA 指纹图谱、进行树种鉴定和遗传相关性的研究。     

杨树重要品种(无性系)的AFLP指纹分析

品种的准确鉴定及其遗传相关性的了解对杨树育种和品种管理具有非常重要的意义。本试验采用AFLP对来自青杨组和黑杨组的21个重要杨树品种（无性系）的鉴定与遗传相关性进行了研究。结果显示,筛选的4对AFLP引物总共产生了181条多态性带,尤其是每对引物对每个品种都产生了独特的指纹图谱;聚类分析和多维尺度分析将试验材料大体上分为五类,结果不仅显示了组间不同品种的差异,而且大体上区分了我国原生品种和外来品种。本研究表明,所有品种都可被筛选的引物准确鉴定,遗传相关性的推断结果与它们的系谱或分类基本一致。另外,本研究还表明AFLP技术完全可用于大规模地构建杨树树种DNA指纹图谱、进行树种鉴定和遗传相关性的研究．     

落叶松转基因育种体系的建立及其产业化应用

落叶松等针叶速生用材树种是北方工业人工林的主体树种,利用传统育种方法对落叶松进行遗传改良已不能满足现代林木遗传育种的需求.同时,由于其基因组庞大（大于15G）,已知基因序列信息不足,成为植物界最难进行细胞工程、染色体工程、基因工程等分子育种及其品种创制的植物种.本文重点介绍了笔者研究组10多年来基于现代基因工程手段建立的农杆菌介导的落叶松遗传转化体系及花粉管介导的遗传转化体系,并介绍了利用转基因技术对落叶松进行快速定向培育及遗传改良的研究成果.     

浙闽五个红豆树自然保留种群的遗传多样性

红豆树是我国三级保护的珍稀濒危植物,是家具、装饰、雕刻等上等用材。应用ISSR分子标记研究浙闽2省5个红豆树自然保留种群的遗传多样性和种群遗传分化。结果表明,红豆树遗传多样性丰富,物种水平的多态位点百分率高达91．46％,总的种群基因多样性为0．3981,显著地高于其他珍稀濒危树种。研究的5个红豆树自然保留种群虽较小,但皆维持较高水平的遗传多样性,种群多态百分率（PPL）、Nei基因多样性（HE）和Shannon信息多样性指数（I）分别为81．71％～89．02％、0．3498—0．3831和0．5026～0．5506。因现有红豆树种群皆是在过渡采伐后保留下来的,片断化时间较短,种群间遗传分化较小,仅有6．45％的遗传变异存在于自然保留种群间,而种群内的变异占总变异的93．55％。较大的红豆树种群遗传多样性相对较高,应优先加强遗传保育。     

树种多样性和土壤微生物多样性对人工林生产力的影响

林分生产力通常会随着树种多样性增加而增加,但不同营养级生物多样性以及树种和土壤微生物多样性之间的相互作用如何影响生产力目前尚不清楚。以亚热带不同丰富度和树种组成的人工林为研究对象,从物种、功能性状、遗传三个维度的树种多样性以及土壤真菌和细菌系统发育多样性,探究了中国亚热带人工林树种多样性和土壤微生物多样性对林分生产力的影响。研究发现,林分生产力随树种功能多样性(FD)(P<0.001)、比叶面积群落加权均值(CWM-SLA)(P<0.01)、树种系统发育多样性(PD)(P<0.05)和土壤真菌多样性(PDF)(P<0.01)的增加而显著增加,分别解释了林分生产力总变异的12.86%、6.80%、3.67%和3.08%。FD和CWM-SLA可分别通过增加土壤真菌、细菌多样性而间接提高林分生产力。研究结果表明多营养级生物多样性是维持高水平林分生产力的基础,树种多样性和土壤微生物多样性之间的自上而下的级联效应在调节生态系统生产力方面发挥着重要作用。     

应用SRAP标记分析新疆地区主要杨属树种的遗传多样性

用30对SRAP引物对15个采集于新疆不同地区的杨属树种进行了遗传多样性分析,共扩增出881条具多态性的条带,平均多态性信息量为0．958,平均多态性比率为96．8％．通过聚类分析将15个杨树材料分为4个群．这一聚类结果与传统的分类基本上一致。这表明SRAP标记适用于研究杨属树种遗传多样性。     

新疆雪莲体细胞胚胎发生

通过体细胞胚胎发生途径实现了新疆雪莲（Saussurea involucrata Kar．et Kir．）的植株再生。选用新疆雪莲子叶为外植体,接种于MS＋0．5mg·L^-12,4-D＋0．05—1mg·L^-1BA的固体培养基上,进行愈伤组织的诱导。从第1次继代培养的愈伤组织中挑选出黄绿色、颗粒状、质地致密的腔陛愈伤组织,转移到含0．05—0．1mg·L^-1 2,4-D的MS液体培养基中进行悬浮培养,20天后可分化产生大量球形胚。继代过程中相继加入PEG和GA3,可以促进体细胞胚的分化和生长。体细胞胚在含有5mg·L^-1 GA3的MS固体培养基上,可发育成完整的植株。     

新疆雪莲体细胞胚胎发生

通过体细胞胚胎发生途径实现了新疆雪莲(Saussurea involucrata Kar.et Ki r.)的植株再生。选用新疆雪莲子叶为外植体, 接种于MS+0.5 mg.L-1 2,4-D+0.05-1 mg.L-1 BA的固体培养基上, 进行愈伤组织的诱导。从第1次继代培养的愈伤组织中挑选出黄绿色、颗粒状、质地致密的胚性愈伤组织, 转移到含0.05-0.1 mg.L-1 2,4-D 的MS液体培养基中进行悬浮培养,20天后可分化产生大量球形胚。继代过程中相继加入PEG和GA3 , 可以促进体细胞胚的分化和生长。体细胞胚在含有5 mg.L-1 GA3 的MS固体培养基上, 可发育成完整的植株。     

Plant Regeneration from Cell Supension Derived Protoplasts of Heracleum moellendorffii

Heracleum moellendorffiz Hance is a herb belonging to Umbelliferae used in traditional medicine in China. The young stem-nodes were induced for callus formation on MS medium containing 1 mg/L 2,4-D. After subcultured for about five months, the embryogenic calli were used for cell suspension culture. The protoplasts were prepared from this suspension by digestion with enzyme mixture containing 1. 5% cellulase Onozuka R-10 +0. 3% macerozyme R-10 + 0. 5% snailase + 5 mmol CaCl2 + 0. 6 mol/L mannitol, at pH 5.8, and cultured in modified MS and modified N6 media with 0.3 % agarose. They divided after 3 days and developed into small cell colonies after about 2 weeks. From this time on, the glucose concentration in the culture media was decreased to 0. 2 mol/L,which led to futher growth of the colonies to small calf . After a period of proliferation on solid medium with 0. 5 mg/L 2,4-D, the calli were transferred to a medium with 0. 1 mg/L zeatin on which somatic embryos differentiated and developed to plantlets     

Plant regeneration through somatic embryogenesis from suspension cultures of a minor millet,Paspalum scrobiculatum

Compact, friable and embryogenic calli were initiated from immature inflorescences and young leaf bases of one week old seedlings of Paspalum scrobiculatum cultured on MS medium supplemented with 2,4-D. A stable, embryogenic suspension culture was initiated from these calli and maintained in a liquid version of the same MS medium. Embryogenic calli and somatic embryos were obtained by plating suspension culture cells onto semi-solid medium containing 2,4-D. Complete, normal plantlets developed on 2,4-D free medium at a high frequency from somatic embryos. NAA and BAP in the medium promoted plant development.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - BAP 6-benzylaminopurine - ABA Abscisic acid - MS Murashige and Skoog (1962) - CM Coconut milk     

云南大叶茶体细胞胚发生及体细胞胚苗形成体系的建立

利用云南大叶茶(Camellia sinensis var.assamica Kitamura)胚性细胞系(CL_1)中悬浮培养物,建立了高频率同步化体细胞胚发生及体胚苗形成体系。以改良的MS为基本培养基,将CL_1中培养物由液体保持培养基(0.1mg/L 2,4-D 0.5mg/L 6-BA)继代转入液体诱导培养基(0.05mg/L 2,4-D 0.50mg/L6-BA),暗培养诱导28d,转入不含任何激素的液体分化培养基中再培养28d,获得了不同发育时期的体细胞胚,其发生频率为81.5％。不同发育时期的体细胞胚用不同目的细胞筛收集,在液体生长培养基(1/2 MS 1.0mg/L GA_3 0.5mg/L 6-BA)中培养发育成熟。ABA有利于高质量体细胞胚的形成。20～70月大小的体细胞胚在固体生长培养基中成苗转换率为75％。在液体悬浮培养条件下观察记录了体细胞胚发育过程,证实其过程与合子胚的形态发生过程相似。     

新疆天山雪莲体胚诱导与分化研究

以新疆天山雪莲的叶片为外植体,分别用不同配方培养基诱导愈伤组织,后进行体胚诱导和分化培养形成再生雪莲植株.结果表明,诱导愈伤组织的最适培养基为MS 2,4-D 0.5 mg/L BA 1.5 mg/L,诱导率可达到100%;愈伤组织转移至MS 2,4-D 0.5 mg/L BA 1.5 mg/L培养基进行继代培养,增殖后的愈伤组织转移到MS 2,4-D 0.2 mg/L的液体培养基后成功诱导出雪莲体胚,出胚率达40%;将体胚接至MS ABA 0.5 mg/L培养基后,结果分化生长出大量的再生雪莲幼苗.     

Somatic embryogenesis and plant regeneration from protoplasts of asparagus (Asparagus officinalis L.)

Protoplasts were isolated from embryogenic calli of Asparagus officinalis L. cv. Mary Washington and cultured in 1/2 MS medium with 1 mg/l NAA, 0.5 mg/l zeatin, 1 g/l L-glutamine, 0.6 M glucose and 0.1% Gellan Gum. Protoplasts started to divide after 3–4 d of culture and formed visible colonies after 30 d of culture. The percentage of colony formation (plating efficiency) was 7.2%. The colonies were then transferred onto Gellan Gum-solidified MS medium containing 1 mg/l 2,4-D and 3% sucrose for further growth. Somatic embryos were induced from all colonies of 0.5–1.0 mm size after transferring to 1/2 MS medium lacking growth regulators. After treating these somatic embryos (1–3 mm) in distilled water for a week, 30–40% of them germinated normally and grew into plantlets 20–30 d after transplanting on 1/2 MS medium containing 1 mg/l IBA, 1 mg/l GA₃ and 1% sucrose. These protoplast-derived plants were diploid with 20 chromosomes.Abbreviations BA 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - GA₃ gibberellic acid - IBA indole-3-butyric acid - MS Murashige and Skoog (1962)     

毛白杨悬浮细胞系的建立及再生植株的获得

以毛白杨基因型TC152无菌苗为材料,研究毛白杨悬浮细胞系建立与植株再生,结果表明,通过悬浮培养和固体培养两种方法诱导毛白杨悬浮细胞分化不定芽,最终获得无菌生根苗。愈伤组织在MS＋1.5mg·L-12,4-D＋30g·L-1蔗糖的液体培养基中振荡培养,12d可建立悬浮细胞系;悬浮细胞系继代培养基为MS＋0.8mg·L-12,4-D＋30g·L-1蔗糖,继代周期为7d,悬浮细胞在MS＋1.0mg·L-16-BA＋0.1mg·L-1NAA＋0.5～1.0mg·L-1ZT＋30g·L-1蔗糖培养基中悬浮培养,可分化大量不定芽,每个培养瓶中可得到40～50个芽,个别不定芽玻璃化;不定芽在1/2MS＋0.6mg·L-1IBA＋20g·L-1蔗糖＋5.5g·L-1琼脂培养基上可分化不定根。悬浮细胞通过固体平板培养增殖为愈伤组织块后,在MS＋1.0mg·L-16-BA＋0.1mg·L-1NAA＋1.0mg·L-1ZT＋30g·L-1蔗糖＋5g·L-1琼脂的固体培养基上,不定芽分化率可达到70.00%。     

Establishment of a System with High Synchronous Frequency of Somatic Embryogenesis and Embryo Seedling Formation in Camellia sinensis var. assamica

The system of high synchronous frequency of somatic embryogenesis and somatic embryo seedling formation was established by means of embryonic cell hne 1 ( CL1 ) of Camellia sinensis var. assamica Kitamura. Modified MS was used as the basic medium. Cultures of CL1 was transferred to the aqueous induced medium (0.05 mg/L 2,4-D + 0.50 mg/L 6-BA) from the maintenance medium (0.1 mg/L 2,4-D + 0.5 mg/L 6-BA) for somatic embryos induction under dark condition. 28 days later, they were cultured in the liquid differentiation medium. Various kinds of somatic embryos were obtained after another 28 days. The frequency of somatic embryos was 81.5 %. Various mesh sizes of sieves were applied to collect the somatic embryos in different developmental stages which could develop to mature stage in the aqueous growth medium ( 1/2 MS + 1.0 mg/L GA3 + 0.5 mg/L 6-BA). ABA was effective to promote the formation of highly qualified somatic embryo. The mature somatic embryos sized 20 to 70 mesh had the conversion frequency 75 %. The development of somatic embryogenesis studied under a cell suspension culture system was similar to the zygotic embryogenesis.     

Plant regeneration via somatic embryogenesis from solid and suspension cultures of Vitis vinifera L. cv. ‘Manicure Finger’

Vitis vinifera L. cv. ‘Manicure Finger’ is one of the major table grape varieties in China. To provide a strong foundation for genetic transformation with potential for crop improvement, we undertook plant regeneration via somatic embryogenesis. Anthers and gynoecia were harvested from immature flowers and used as explants to induce embryogenic calli. Explants cultured in MS1 medium (based on Murashige and Skoog basal salts), supplemented with 4.5-μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.4-μM 6-benzylaminopurine (6-BA) showed the highest rates of embryogenic callus induction (3.7%?±?1.3% for anthers and 4.8%?±?2.5% for gynoecia). After several months, somatic embryos were produced from embryogenic calli cultured in plant growth regulator-free MS2 medium (with reduced sucrose). Somatic embryos (SE) at the cotyledonary stage were isolated and cultured on three different media (MS2, MS3, or B) for conversion into plantlets, the efficiency of which ranged from 63.9%?±?4.8% to 83.9%?±?8.4%. After 1 mo of in vitro culture, 80% of plants with at least six leaves were successfully transplanted into soil. SE was repeatedly induced from previously induced somatic embryos for up to 1.5 yr. Using embryogenic calli as starting material, suspension cultures containing embryogenic cell aggregates were also established in liquid MS medium supplemented with 4.5-μM 2,4-D. The embryogenic cell aggregates continued to proliferate without differentiating for successive subculture cycles. After transfer to 2,4-D-free liquid medium for 4 wk, an average of 63.7%?±?9.0% mature SEs were produced per 20 mL of liquid medium. More than 40% of somatic embryos at cotyledonary stage, derived from the suspension cultures, successfully germinated into plants using solid medium.