荔枝愈伤组织诱导及其体细胞胚胎发生研究初报

分别取元红荔枝盛花后45d未败育的幼胚、一个月龄的实生苗叶片、成龄树叶片、未受精子房接种在愈伤组织诱导培养基上,均能诱导出愈伤组织,但仅有幼胚能诱导出胚性愈伤组织。幼胚在含5% 蔗糖的MS基本培养基附加2,4-D2mg/L+水解乳蛋白500mg/L的培养基上诱导率最高。幼胚愈伤组织在含5% 蔗糖的基本培养基附加10% 椰乳的培养基上,白色胚状体发生率最高。胚状体易提前萌发,仅抽生根不抽生茎。健壮的未生根胚状体在含5% 蔗糖的培养基附加BA1mg/L的培养基上生根率最低,为理想的萌发培养基。随着BA浓度的降低,生根率提高,呈负相关。同浓度BA抑制生根的效果好于KT。在萌发培养基上生长一段时期后,少数较成熟的胚状体抽生出茎叶,长成完整植株。     

3种东方百合离体培养

【目的】研究3种东方百合离体培养技术,筛选出适宜其离体培养的统一培养基,为东方百合工厂化育苗提供技术保障。【方法】以3种东方百合提伯、索邦和西伯利亚的鳞片为外植体,筛选适宜的灭菌方法;在MS基本培养基上添加不同种类和不同浓度的激素,筛选适宜的增殖培养基和生根培养基,进一步筛选最适移栽基质。【结果】适宜3种东方百合外植体灭菌的较好方法是:75%酒精灭菌15 s结合升汞灭菌12 min;愈伤诱导和分化的适宜培养基为MS;MS+0.5 mg/L 6-BA+0.5 mg/L NAA是增殖培养的最适培养基,提伯、索邦和西伯利亚的增殖系数分别为3.32、3.32和3.00;MS+0.1 mg/L IBA是生根培养的最佳培养基,提伯、索邦和西伯利亚的生根率分别为96.33%、92.67%和93.67%;移栽的最佳基质为泥炭∶珍珠岩(体积比)=2∶1,提伯、索邦和西伯利亚的移栽成活率分别为97.34%、98.00%和100.00%。【结论】建立的3种东方百合离体快繁体系适用于东方百合种球工厂化生产。     

‘夏日阳光’番茄侧芽离体培养快繁技术

‘夏日阳光’番茄是近年来引进的高端品种,丰产性好、品质高,深受市场的青睐,但其种子为F1代杂种,价格昂贵,很难在市场上大面积推广。利用植物离体培养技术快速繁殖‘夏日阳光’番茄种苗,是生产上推广该品种的一条可行途径。为快速建立‘夏日阳光’番茄无菌株系以及获得其试管芽苗的高效增殖,以‘夏日阳光’番茄侧芽为材料,探讨其离体培养过程中的关键技术。试验结果表明,采用0.1%Hg Cl2溶液(w/v)加2滴土温20消毒8 min的处理方式消毒后,培养在MS+0.1 mg/L BA+0.1 mg/L IBA培养基中,可以较快速地建立‘夏日阳光’番茄无菌株系,成苗率为46.7%;继代增殖适宜的培养基配方为MS+0.5 mg/L BA+0.1 mg/L IBA,增殖系数可达4.3,并且芽苗生长健壮,为其种苗的规模化繁育提供技术基础。     

刺山柑腋芽的组织培养快繁技术研究

以从意大利西西里岛引进的刺山柑带腋芽茎段为外植体,进行了预处理、丛芽诱导、丛芽增殖和生根、再生植株炼苗等研究,采用正交设计筛选出各阶段的适宜培养基.结果表明:在诱导阶段,较低的激素水平能促进幼芽萌发,其适宜的培养基为DKW+0.3 mg·L-16-BA+0.05 nag·L-1NAA+20 g·L-1庶糖;继代增殖阶段,丛芽增殖系数与6-BA用量成正比,但随着6-BA用量加大或长期在高浓度6-BA上培养,芽苗多且细弱,不适合做生根诱导,其适宜的培养基为DKW+0.3 mg·L-16-BA+0.025 mg·L-1NAA+30 g·L-1庶糖;生根培养阶段,其适宜的培养基为1/2MS+0.2 nag·L-1 IBA+100～200 mg·L-1 VB1+20 g·L-1庶糖,生根率87%.生根组培苗移植至草炭灰:珍珠岩(3:1)基质中成活率为92.31%.     

鳞片不同放置方式对卷丹快速繁殖的影响

以不同放置方式的卷丹鳞片为外植体,进行组织培养已成功获得再生植株,并已建立快速无性繁殖体系.3种不同放置方式分别为:鳞片内侧向上平放、鳞片竖直插入、鳞片内侧向下平放.以MS培养基+不同的激素配比进行培养,结果表明,不同放置方式的鳞片分化能力由强到弱依次为:鳞片内侧向上平放＞鳞片竖直插入＞鳞片内侧向下平放.     

膏桐组织培养的初步研究

 通过不同激素配比诱导膏桐外植体产生愈伤组织,为利用愈伤组织进行离体快繁和膏桐转基因及生物工程育种奠定基础。结果表明：诱导膏桐子叶产生愈伤组织的最佳培养基是MS+2,4-D 2.0mg/L和 MS+6-BA 2.0mg/L+NAA 2.0 mg/L;诱导愈伤组织形成芽的最佳培养基为：MS+6-BA 2.5 mg/L+IBA 0.1mg/L;根的形成不需添加任何激素。适当的激素配比可有效诱导膏桐子叶产生愈伤组织并形成再生植株.     

Large scale in vitro propagation of Stevia rebaudiana (bert) for commercial application: Pharmaceutically important and antidiabetic medicinal herb

Stevia rebaudiana is a valuable medicinal plant species and it is being used for the treatment of diabetes. Currently, there is a high demand for raw material of this medicinal herb due to ever increasing diabetes disorder among the population. In order to meet the increased demand an efficient in vitro propagation of S. rebaudiana was established. Nodal explants collected from the field were cultured on MS basal medium fortified with different concentrations of BAP (0.5-3.0 mg/l) and KIN (0.5-3.0 mg/l) individually for shoot bud induction. In vitro derived nodal buds were cultured on MS medium supplemented with different concentrations (0.5-3.0 mg/l) of BAP and KIN for multiple shoot bud regeneration. In the second experiment, in vitro derived buds were placed on MS medium supplemented with different concentrations of BAP (0.5-3.0 mg/l) in combination with 0.5 mg/l IAA or IBA or NAA for shoot bud multiplication. The highest frequency (94.50%) of multiple shoot regeneration with maximum number of shoots (15.69 shoots/explant) was noticed on MS medium supplemented with 1.0 mg/l BAP. For large scale plant production, in vitro derived nodal bud explants were cultured on MS medium fortified with 1.0 mg/l BAP, in which about 123 shoots/explant were obtained after three subcultures on the same media composition. Elongated shoots (>2 cm) dissected out from the in vitro proliferated shoot clumps were cultured on half-strength MS medium containing different concentrations of NAA (0.1-0.5 mg/l) and/or MS medium fortified with various concentrations (0.5-2.0 mg/l) of auxins (NAA, IAA and IBA) for root induction. Highest frequency of rooting (96%) was noticed on half-strength MS medium augmented with 0.4 mg/l NAA. The rooted plantlets were successfully transferred into plastic cups containing sand and soil in the ratio of 1:2 and subsequently established in the greenhouse. The present in vitro propagation protocol would facilitate an alternative method for rapid and large-scale production of this important antidiabetic medicinal plant.     

植物组织培养中污染控制技术的研究现状

分析了植物组织培养过程中的污染原因,结合外植体消毒技术的研究现状,提出如何控制由外植体引发污染的方法。选择合适的外植体和消毒剂,采用联合消毒方法,对外植体初代培养中内源污染的控制效果比较明显。另外,对于难消毒的外植体,配合使用抗生素能有效地控制污染。     

荔波野生金线莲芽的诱导及其增殖

为探寻金线莲组织快繁的配套技术,以金线莲茎段、茎尖为外植体,研究NAA、6-BA浓度梯度及配比、不同部位外植体对不定芽诱导和增殖的影响.结果表明:MS+6-BA 1.0 mg/L+NAA 0.1 mg/L为适宜的诱导芽和增殖培养基,培养50 d后增值系数达6.5,MS+6-BA 1.0 mg/L+NAA 0.15 mg...     

文冠果不定芽再生与快速繁殖

以成熟文冠果叶片作为外植体,通过器官直接发生途径,对其进行不定芽诱导、不定芽增殖及生根培养,建立了一种简单易行的、可再生的文冠果快速繁殖方法。结果表明,在不定芽诱导阶段,当MS培养基添加2.0mg.L-1BA和0.2mg.L-1 NAA时,不定芽诱导率最高;高浓度的BA显著降低不定芽诱导率;当MS培养基添加2.0mg.L-1 BA和0.1mg.L-1 NAA,每个外植体获得的不定芽数达到最大;当叶片远轴面接触培养基时,不定芽诱导率高于近轴面接触培养基。在增殖培养阶段,当MS培养基添加0.5mg.L-1 BA时,不定芽增殖率、增殖个数及增殖芽长均达到最大;在不同浓度的BA培养基中添加NAA降低了不定芽的增殖效果。在生根培养阶段,1/2MS培养基添加0.5mg.L-1 IBA的生根率最高;IBA的生根效果显著优于NAA或IAA。生根苗在含有土壤、沙子和泥炭的混合基质(1∶1∶1)中移栽成活。