多年生黑麦草高频再生体系的建立及农杆菌介导PEPC基因的转化

以多年生黑麦草(Lolium perenne L.)成熟种子作为外植体,建立其高频再生体系,通过农杆菌介导法将磷酸烯醇式丙酮酸羧化酶(PEPC)基因转入,为多年生黑麦草的抗逆转基因工作奠定基础。结果表明:种子充分灭菌后在附加6.0 mg·L^-1 2,4-D的培养基中诱导愈伤组织,诱导率可达61.33%。愈伤组织在2,4-D浓度减半的培养基上继代培养长势良好,分化培养基为MS培养基附加1.0 mg·L^-16-BA,分化率最高达到61.33%,在1/2 MS+0.5 mg·L^-1 NAA培养基中进行生根培养,生根率达100%。以该再生体系为基础,将获得的愈伤组织作为外植体进行遗传转化,经筛选培养后得到再生植株,通过PCR及实时荧光定量(Real-Time)PCR验证表明PEPC基因已成功转入,转化率为8.67%。试验结果将为多年生黑麦草抗性品种的研究奠定基础。     

葡萄组织培养中愈伤组织的诱导

采用三因素(萘乙酸NAA,吲哚丁酸IBA,6-苄氨基嘌呤6-BA)四水平的正交试验设计,对"鄞红"葡萄近梢处的嫩茎段和幼果进行愈伤组织诱导试验。结果表明,6-BA对愈伤组织诱导影响最大。以嫩茎段为外植体的最佳培养基配方是MS+1.00 mg/L 6-BA+0.10 mg/L NAA+0.20 mg/L IBA,其诱导率为70.00%。以幼果为外植体的最佳培养基配方是MS+0.80 mg/L 6-BA+0.10 mg/L NAA+0.20 mg/L IBA,诱导率为61.85%。嫩茎段是"鄞红"葡萄愈伤组织诱导最理想的外植体。     

紫外线诱变处理对苜蓿种子苜蓿皂苷含量的影响研究

以紫外线诱变处理苜蓿种子,接种在Ms培养基诱导产生愈伤组织,用微波辅助提取的方法提取其中的皂苷类物质并测定其中的皂苷含量。结果表明：诱变处理苜蓿种子产生的愈伤组织的皂苷含量高于未处理的种子0.3％,紫外线诱变可提高苜蓿皂苷含量。     

Factors influencing mansonone induction in elm callus by culture filtrate elicitors of Qphiostoma ulmi

Mansonone F (phytoalexin) production in elm callus induced by elicitors of Ophiostoma ulmi is affLited by cultural and experimental conditions. These factors include different fun gal isolates, fungal culture age, elicitor concentration, the time of elicitor treatment, the callus cu Ytured with or without light and the callus subculturing frequency.     

紫花苜蓿胚性愈伤组织诱导与增殖的研究

为在不同栽培苜蓿(Medicago sativa)品种上,建立稳定、快速且高效的组织培养体系,本试验以品种'游客’、'Paola’和'Sardi7’为研究材料,探究了苜蓿外植体、激素配比及金属离子(Cu²⁺、Ag⁺),对其胚性愈伤组织的诱导效果。结果表明：下胚轴为诱导愈伤组织的最佳外植体;MS+1 mg·L^-12,4-D+0.2 mg·L^-1 KT为培养基时'游客’和'Paola’胚性愈伤诱导率分别为24.7%,16.7%;MS+2 mg·L^-12,4-D+0.2 mg·L^-1 KT为培养基时'Sardi7’胚性愈伤诱导效果最佳为27.3%;1 μmol·L^-1 和5 μmol·L^-1 Cu²⁺均能显著促进'Sardi7’'游客’和'Paola’愈伤组织的诱导;5 μmol·L^-1 Ag⁺显著诱导'游客’和'Paola’形成胚性愈伤组织,诱导率分别提高25.3%,46.7%(P<0.05)。本研究探索了诱导苜蓿愈伤组织的最佳外植体及其培养条件、金属离子促进不同苜蓿品种再生的作用,为突破同源四倍体栽培苜蓿品种基因型上的限制、提高胚性愈伤组织出愈率提供了思路。     

苹果砧木M26离体叶片愈伤组织发生及分类研究

对苹果砧木Ｍ２６ 离体叶片愈伤组织的发生研究表明,取继代后３０ ～３５ ｄ 的试管苗顶部１～４ 叶位的叶片,沿与主脉垂直方向横切后,以叶背面朝上方式置于愈伤组织诱导培养基上,经观察在叶片近柄端切口处愈伤组织发生较早,产生量较多。最适的愈伤组织诱导培养基为：ＭＳ＋２ ,４ － Ｄ０ ．２ ～２ ．０ ｍｇ·ｌ－ １ ＋ＩＡＡ０ ．２ ～２．０ ｍｇ·ｌ－ １ ＋ ＢＡ０ ．５ ～１．０ ｍｇ·ｌ－ １ ＋ 蔗糖３ ％ ＋ 琼脂０．６％ 。将诱导产生的愈伤组织按外部形态、细胞特点及生理生化指标等分为三类,其中表面干燥致密、细胞内含物丰富、淀粉、蛋白质及干物质含量均较高的Ⅲ类愈伤组织的芽再生频率最高,可达８３ ．３３ ％ ,是能利用的愈伤组织类型。     

石海椒愈伤组织诱导与增殖研究

对石海椒幼嫩叶片和茎段进行了愈伤组织的诱导增殖研究,以期探寻其诱导和增殖的最佳条件。结果表明:石海椒的叶片和茎段都能诱导出愈伤组织,但最佳外植体为叶片,叶片和茎段在0.1%升汞中适合消毒时间为6 min和10 min。叶片最适诱导培养基为B5+0.6 mg/L6-BA+2.0 mg/L NAA,增殖培养基为B5+0.6 mg/L 6-BA+1.25 mg/L NAA。基本培养基和生长调节剂对愈伤诱导的影响是NAA>6-BA>基本培养基。     

Rejuvenation influences indirect organogenesis from leaf explants of Pomegranate (Punica granatum L.) ‘Kandhari Kabuli’

Sequential subculturing leads to a gradual physiological change in cells that may be termed ‘rejuvenation’. The effect of repetitive subculturing on callus induction and shoot regeneration from leaf explants of Punica granatum L. ‘Kandhari Kabuli’ were investigated. Surface-sterilised leaves were cultured on 1.0× Murashige and Skoog (MS) medium supplemented with 4.0 mg l^–1 α-naphthaleneacetic acid (NAA) and 2.0 mg l^–1 6-benzyladenine (BA) for callus induction. Shoots were regenerated from callus on 1.0× MS medium supplemented with 1.5 mg l^–1 BA, 0.5 mg l^–1 kinetin, and 0.25 mg l^–1 NAA. Subculturing of callus onto fresh medium maintained the rate of shoot formation and substantially increased the production of shoot buds up to the second subculture. Following further subculture passages, a lower shoot regeneration potential from callus was observed. A maximum shoot bud induction from callus of 63.9% was observed at the second subculture passage. The rate of multiplication of in vitro shoots increased until the fourth subculture, then became constant. Similarly, in vitro rooting of micro-shoots increased up to the third subculture, followed by a decline during further subculturing.     

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     

玉米叶片胚性愈伤组织诱导及其与内源IAA和ABA关系的初步研究

玉米幼叶愈伤组织的形成能力因其形态学部位的不同而呈显著差异, 从基部向上依次递减, 幼叶外植体的内源IAA和ABA水平亦相应地呈递减趋势。 2, 4-D及与BA组合对基部切段的愈伤组织形成无显著影响, 愈伤组织生长符合Logistic曲线, 单独使用BA不利于愈伤组织形成; 采用2 mg/L 2, 4-D与0.5 mg/L BA的组合最有利于胚性愈伤