转rol基因枳橙分子鉴定及部分生物学的观测

 对转rol基因枳橙B、D、E 3个系及对照田间高接植株进行了PCR分子鉴定、RT-PCR转基因表达分析和生物学特征的观测。PCR分析表明rolA、rolB、rolC已成功转入到3个转化枳橙系中。利用RT-PCR在田间转化植株中只检测到nptⅡ和rolC基因的表达, 而rolA、rolB基因未检测到。3个转化系枳橙均表现出顶端优势减弱, 侧枝增多, 植株矮化性状明显, 高度只有对照植株的44% ～50% , 节间长度缩短,叶面积减少; 光合作用增强; 内源赤霉素、生长素、玉米素等促进类激素在中部、下部叶片含量均高于对照, 这可能是诱导上述形态特征的变化的因素之一。     

枳橙秋播快速繁育技术

枳橙是甜橙和部分宽皮柑桔的重要砧木，在美国、南非、西班牙和澳大利亚等国被广泛应用。我国从上世纪末开始批量性引进进行容器育苗，目前还处于示范推广阶段，因此，有关枳橙的播种繁育技术少有报道。为此，笔者进行了本试验。     

旱金莲的组培快繁技术

简要介绍了旱金莲的形态特征和生物学习性,重点从材料的选取、接种、继代培养、生根培养和驯化移栽等方面进行论述,为组培快速繁殖旱金莲提供依据。     

千层金组培快繁技术研究

以嫩茎为外殖体,以MS为基本培养基,分别加入不同浓度6-BA、NAA、IBA,研究不同浓度激素及组合对千层金组培快繁的影响。结果表明:诱导芽的最佳培养基为MS+6-BA2.0 mg/L+NAA 0.2 mg/L,诱导率达63.6%;继代培养最佳培养基为MS+6-BA 1.0 mg/L+NAA 0.2 mg/L,增殖系数达6.3,且芽苗健壮;壮苗培养的最佳培养基为MS+NAA 0.1 mg/L,每瓶产苗数达16.2株;最佳生根培养基为1/2MS+NAA 0.1 mg/L,21 d生根率达100%,平均根数达9.6条。采用泥炭土与椰糠体积各半混合基质进行移栽,30 d成活率达95.7%,能够满足工厂化苗木生产的要求。     

树莓组织培养和快繁研究

树莓 ,又称黑莓 ,为蔷薇科悬钩子属植物。具有适应性强 ,果实营养丰富 ,产量高等优点。近年来发展很快 ,但主要采用扦插、压条和分根蘖等方法繁殖 ,既受季节影响 ,又受材料限制 ,造成病虫害传播蔓延 ,带来不必要损失。本文就组织培养快繁生产无病树莓苗木的有关技术问题进行探讨。1　材料与方法　　材料来自福州招宝农业开发公司开发基地 1年生树莓嫩枝。无菌株系建立 :从健康嫩枝上取茎尖部分用自来水冲洗 10分钟 ,在 70 %酒精浸 2 0秒钟 ,然后在0 1%升汞中消毒 10分钟 ,最后用无菌水浸洗 4～ 5遍。消毒处理后的茎尖接种在含有不同植物生…     

红叶石楠微体快繁技术研究

以红叶石楠茎段为材料,采用组织培养方法,研究了红叶石楠的微体快速繁殖技术。结果表明:当年生的半木质化的嫩梢用0.1%的升汞溶液消毒7～8min效果最好;嫩茎启动培养基采用MS+1.5mg/L 6-BA+0.5mg/L KT+0.2mg/L NAA效果最好;增殖分化的培养基为MS+2.0mg/L 6-BA+2.0mg/L KT+0.1mg/L NAA,增殖率可达4.3倍;生根培养基选用1/2 MS+0.5mg/L NAA+1.0mg/L IBA效果最佳。组培苗培养条件为:光照强度1500～2000Lx,光照时间13h/d,培养室温度为23℃。     

大花蕙兰组培快繁技术研究

以大花蕙兰茎尖为外植体,进行组织培养,结果表明,生长点可诱导形成愈伤组织及再生植株.经试验筛选出各培养阶段最适宜的培养基为:愈伤组织诱导培养基:MS BA1.5～2 mg/L;分化培养基:MS BA1.5mg/L NAA 0.5mg/L;生根培养基:1/2MS(大量元素减半) NAA1.5mg/L 0.5%活性炭.     

苹果枣组培快繁技术研究

以MS为基本培养基 ,取苹果枣萌蘖苗茎段作外植体进行离体快繁研究 ,优选确定了其最佳分化、生根培养基的激素组成及各激素的比例关系 ,并且成功地进行了试管苗的移栽。试管苗繁殖系数达 6 2左右 ,生根率达 91 6 % ,移栽成活率达 98%     

紫苏的组织培养及快繁技术研究

以紫苏(Perilla frutescens(Linn)Britt)的种子获得无菌苗,取无菌苗的带节茎段为外植体,对影响紫苏丛芽增殖和生根的主要因子进行了研究.结果表明:紫苏种子较好的灭菌方法为:75%酒精20 s 0.1%升汞8 min;增殖培养基为:MS 6-BA 2.0 mg/L NAA 0.1 mg/L;生根培养基为:1/2 MS 6-BA 1.0 mg/L NAA 0.1 mg/L;且移栽后成活率可达90%以上.     

利用组培技术快繁艾西丝南瓜良种

利用组织培养方法研究艾西丝南瓜适于生产上应用的种苗快繁技术。试验结果：艾西丝南瓜分人培养基以MS BA1、0mg/L IAA0.1-0.5mg/L为宜;适宜于试管苗生根的培养基为1／2MS0;适宜的试管苗移栽驯化基质为细沙、腐叶土、粗沙分层基质。用组培手段培育的试管苗种性不变异,生长、结果习性优于种子苗。     

In vitro propagation of ‘Troyer’ citrange from epicotyl segments

Isolated epicotyl, root meristem and root segment tissues of ‘Troyer’ citrange [Poncirus trifoliata (L.) Rat. × Citrus sinensis (L.) Osbeck] were established in continuous culture to compare their regeneration potential. Callus was obtained from these explants on a Murashige—Skoog (MS) medium containing NAA (10 mg l^?1) and BAP (0.1–10 mg l^?1). Formation of shoots from root segments was direct without callus formation on MS medium containing BAP (10 mg l^?1) and NAA (1 mg l^?1). Shoot formation from epicotyl callus occurred on MS medium containing 0.25 mg l^?1 BAP and 0.1 mg l^?1 NAA. Formation of shoots from epicotyl segments occurred on MS medium containing BAP (0.5 mg l^?1) and NAA (0.1–1.0 mg l^?1), while rooting of regenerated shoots occurred in treatments containing 2.0 mg l^?1 NAA alone. This system provides a rapid method for propagation of ‘Troyer’ citrange.     

In vitro propagation of wavy-leaved Indian paintbrush (Castilleja applegatei Fern.)

Castilleja spp. (Indian paintbrush, Orobanchaceae) are desirable ornamental plants with showy floral bracts that are native throughout much of the western U.S. Propagation of these hemiparasites by seed is usually successful only when a host species is present, and asexual propagation through traditional methods has proven to be extremely difficult. In this study we present an effective shoot culture micropropagation system for Castilleja applegatei Fern., the wavy-leaved Indian paintbrush. In vitro shoot tips of three C. applegatei clones were cultured for 28 days on woody plant medium (WPM) supplemented with four concentrations of each of three cytokinins: 6-benzylaminopurine (BA), 6-(y,y-dimethylallylamino)-purine (2iP) and zeatin. Responding explants, shoot number per responding explant and shoot length were determined. In vitro rooting of microcuttings cultured for 42 days on media containing three concentrations of indole-3-butyric acid (IBA) were also evaluated. A high percent response to shoot induction was observed across all treatments, ranging from 94.5 to 100%. C. applegatei responded best to zeatin, which resulted in both the highest mean shoot number and mean shoot length among the cytokinins tested. The best overall shoot multiplication occurred on media with 4.0 μM zeatin, yielding a mean shoot number of 4.11 and mean shoot length of 3.95 cm. The mean highest rooting response (66.7%), root number (13.21) and root length (2.73 cm) were obtained on WPM supplemented with 10 μM IBA. Significant clone × treatment interactions existed for all variables except mean root number, thus the optimum treatments for both shoot multiplication and root induction were clone-dependent. Rooted microcuttings were acclimated ex vitro with an average success rate of 81.2%. This study demonstrates the considerable potential of an in vitro shoot culture system for the asexual propagation of Castilleja spp.     

非洲菊试管苗叶片的组培快繁

 用非洲菊试管苗叶片作外植体获得了有再分化能力的愈伤组织和正常的再生植株。在适宜培养基上叶片切块的出愈率和出芽率最高可达96 %和90 %; 小苗4 周增殖倍数为10. 85 , 生根率99 % , 最快4周就能从叶片切块成苗。试管苗叶片快繁最适培养基为: 起始MS + 62BA 3. 0 mg·L^-1 + NAA 0. 1 mg·L^-1; 增殖MS + 62BA 3. 0 mg·L^-1 + NAA 0. 2 mg·L^-1; 生根1/ 2 MS + IAA 1. 0 mg·L^-1 。用作外植体的试管苗以3～4叶期为佳, 叶片切块以4 mm ×4 mm为宜。     

内39号葡萄株系的离体快繁技术研究

利用一个新育鲜食葡萄内39号株系,在原离体快繁技术基础上,改进取材时间和繁殖技术,同时配合试管苗绿枝扦插,经过9个月的繁殖,生产出2.32万株生产用苗。逐月统计了实际生产数,表明葡萄离体快繁技术确实可缩短育种年限,用于育种实践。     

Clonal propagation of mulberry (Morus indica L. cultivar M-5) through in vitro culture of nodal explants

A high frequency of sprouting (80.0%) and shoot differentiation was observed in the primary cultures of nodal explants of Morus indica L. cultivar M-5 on MS medium supplemented with 2,4-D (0.3 mg/l). In vitro proliferated shoots were multiplied rapidly by culture of shoot tips on MS medium with BAP (0.5 and 1.0 mg/l) which produced the greatest multiple shoot formation. Multiplication was also achieved by culture of shoot tips on MS medium with BAP (4.0 mg/l) and GA₃ (0.05 mg/l) which facilitated the elongation of shoots followed by sprouting of axillary buds of in vitro grown shoots. A high frequency of rooting (86.7%) with development of healthy roots was observed from shoots cultured on medium with 2,4-D (1.0 mg/l). Plants with well developed roots were transferred to soil with a survival frequency of 80%.     

枣茎段组织培养的研究

以5个枣品种的茎段为外植体,进行了不定芽的诱导和成苗的研究。在培养基MS+ZT1.75mg/L+KT2.0mg/L+NAA穴0～1.0mg/L雪上,枣茎段可以直接分化不定芽,MS+ZT1.75mg/L+KT2.0mg/L+NAA0.015mg/L培养基最适宜茎段不定芽的诱导。5个枣品种在不定芽的诱导方面存在较大差异培养60d时鸡蛋枣的增殖倍数达到3.0,而尖枣仅0.32。保持叶片2～3枚有利于枝条生根,在1/2MS+IBA0.8mg/L培养基上生根率达90%以上。     

合果芋组织培养快繁技术研究

以合果芋幼嫩侧芽为外植体,研究了不同激素浓度对合果芋组织培养的影响.结果表明:用0.1%的氯化汞溶液+3滴Tween20对外植体处理35 min,灭菌效成功率达75%;较好的诱导培养基为MS+BA 5.0 mg/L+IAA 0.2 mg/L,最佳的增殖培养基为BA 2.0 mg/L+NAA 0.1 mg/L,最好的生根培养基为MS+NAA 0.3 mg/L+IAA 0.2 mg/L;生根瓶苗置于室外自然光线下练苗7 d后,移栽到珍珠岩∶腐殖土∶壤土=1∶1∶1的基质中,成活率可达94%以上.     

金叶欧洲山梅花的组织培养及快速繁殖

以金叶欧洲山梅花带腋芽的幼嫩茎段为外植体,进行组织培养研究.结果表明:不同浓度激素组合配比对金叶欧洲山梅花的分化、生根有显著影响.MS 6-BA 1.5 mg/L NAA1.5mg/L为最佳分化培养基;最佳生根培养基为MS NAA0.7mg/L,生根天数为7 d,生根率可达95%;金叶欧洲山梅花生根苗在不同的移栽基质成活率不同,在草炭∶蛭石∶珍珠岩=7∶2∶1的基质中,试管苗的移栽成活率可达93%,可以进行大量生产.     

乙肝表面抗原大蛋白(S1S2S)基因在转基因苹果中表达

构建了乙肝表面抗原大蛋白基因PRS-S1S2S的表达载体pYF9616,通过根癌农杆菌介导法首次将该基因导入苹果品种红爱佳中,得到了抗卡那霉素的GUS阳性转化植株。随机取3株GUS染色阳性的转基因苹果植株经PCR扩增及RT-PCR检测证实该基因已整合入转基因苹果的基因组,并在转录水平得到了表达,ELISA检测证明在苹果植株中正确表达了乙肝表面抗原大蛋白基因。