Effects of Physiological Status of Parent Plants and Culture Medium Composition on the Anther Culture of Sorghum

Abstract

Anthers at various maturation stages were collected from seven genotypes of Sorghum bicolor, and cultured on MS medium supplemented with 3 mg L^?1 2,4-D and 2.0 mg L^?1 kinetin. The rate of callus induction was high in young anthers, and decreased with maturation of the anther. Young anthers were collected from the plants grown in the greenhouse (greenhouse anthers), in the field (field anthers), and in the chambers kept at 20, 25 or 30 °C. Then they were cultured on MS medium containing 2.0 mg L^?1 kinetin, 1.0 mg L^?1 IAA and 2.5 mg L^?1 2,4-D, on which the rate of callus induction was the highest. The average callus-induction percentage was 6.4% in the field anthers, and 3.7% in the greenhouse anthers. Callus induction was low in the anthers collected from the plants grown at 30° C. Among the genotypes examined, TX403 produced calli at the highest frequency. The rate of plant regeneration from anther-calli was the highest on the medium containing 2.5 mg L^?1 kinetin and 3.0 mg L^?1 IAA. The regeneration rate of green plantlets on this medium was 27% on the average, and was 46.2 and 28.6% in PP290 and TX403, respectively. The chromosome number in root-tip cells was unstable, but was 20 in most of the regenerated plantlets.     

基于玉米自交系E28的农杆菌介导与基因枪转化体系的建立及比较

以玉米自交系E28的胚性愈伤组织为受体,分别利用农杆菌介导法和基因枪轰击法对携带有GUS基因和Bar基因的质粒进行转化,借助GUS基因的瞬时表达率和抗性愈伤组织比例建立体系,并对两种方法进行比较。农杆菌介导法以OD6000.6、浸染30 min、25℃共培养3 d、加入0.01%的silwet L-77为最佳参数;基因枪轰击法为4.5 MPa、11 cm轰击距离最佳参数。农杆菌介导法与基因枪轰击法的GUS瞬时表达率没有显著差异,基因枪法的抗性愈伤组织比例显著高于农杆菌介导法,基因枪轰击法较农杆菌介导法更有利于获得转基因后代。     

红掌愈伤组织培养及不定芽分化影响因素的研究

[目的]构建红掌愈伤组织培养再生植株的技术体系.[方法]以红掌叶片和叶柄愈伤组织为继代培养的外植体材料,探讨红掌品种、外植体大小、激素条件对愈伤组织不定芽分化的影响.[结果]红掌品种YNG1叶片愈伤组织的不定芽分化率最高,叶柄愈伤组织的不定芽分化率则以品种YNG2最高,而品种YNG3和YNG5仅叶片愈伤组织能分化不定芽,叶柄愈伤组织均无分化;与细胞分裂素6-BA相比,向分化培养基中添加ZT与TDZ可促进叶片愈伤组织的增殖及不定芽分化;选取直径约1.0～1.5 cm的愈伤组织块进行继代培养,不定芽分化较多,同时可缩短不定芽分化时间.[结论]该研究为构建稳定高效的红掌组培快繁技术体系提供了新的试验依据.     

Ecophysiological aspects of in vitro biotechnological studies using somatic embryogenesis of callus tissue toward protecting forest ecosystems

This review on current biotechnological methods in forestry for in vitro tissue cultures to define the effect of stress conditions on trees,concentrates on somatic embryogenesis.Callus tissue,the key product of somatic embryogenesis,grows over a tree wound under ex vitro conditions.Callus tissue can be used in research in areas such as pathogenic susceptibility at the embryonic level,effect of heavy metals,influence of low temperatures(cryopreservation),production of secondary metabolites and transformation of plants.Callus of arborescent plants can be induced in vitro by fungal elicitors to produce secondary metabolites for pharmaceutical and cosmetic industries and are strongly repellant to herbivores and can thus act to protect forests.Analyses of dual cultures demonstrated that callus tissue exposed to a pathogenic fungus responds by synthesizing low-molecular-mass proteins belonging to an immune protein class.Cryopreservation of embryonic callus tissue also has broad applications,e.g.,for valuable plant genotypes in gene banks.Without strategies to protect forests against stressfactors,forest ecosystems will degrade to the detriment of all life,including humans.In vitro biotechnological research using callus tissue contributes to progress in forestry and the disciplines of ecology,physiology,phytopathology,culture and selection of plants.     

不同浓度生根液对油橄榄扦插生根情况的影响

将佛奥和鄂植两个品种油橄榄扦插条放入生根液浓度分别为0mg·L^-1、100pmm、200mg·L^-1、300mg·L^-1、400mg·L^-1的生根液激素中浸泡,进行油橄榄扦插条生根率的研究。结果表明,没有生根液浸泡的插穗虽有少量的产生愈伤组织,但几乎不会生根;佛奥在0mg·L^-1、100mg·L^-1、200mg·L^-1、300mg·L^-1、400mg·L^-1浓度生根液浸泡下的生根率分别为3%、37%、42%、76%、63%;鄂植在0mg·L^-1、100mg·L^-1、200mg·L^-1、300mg·L^-1、400mg·L^-1浓度生根液浸泡下的生根率分别为2%、21.25%、40.5%、70.25%、53.13%;由此可以得出,油橄榄扦插浸泡生根液激素的最适浓度为300mg·L^-1。     

楸叶泡桐杂种无性系9503的组织培养技术研究

以楸叶泡桐和白花泡桐杂交无性系的枝条为外植体,开展了离体芽增殖技术,以及愈伤组织诱导、芽分化和不定芽生根等组织培养技术的研究。结果表明,增殖培养基采用MS+8mg/L6-BA+0.3mg/LNAA,不定芽的增殖倍数可达到9;利用其叶片进行愈伤组织诱导的最佳培养基为MS+12mg/L6-BA+0.4mg/LNAA,诱导率高达95%;愈伤组织分化的最佳培养基为MS+10mg/L6-BA+0.7mg/LNAA,不定芽生根的最佳培养基为1/2MS+0.5mg/LIBA+0.3mg/LNAA,生根率100%。     

‘新海16’愈伤组织诱导及白霜的优化

为获得高品质、活力旺盛且具有分化能力的愈伤组织,本研究在已建立的海岛棉(Gossypium barbadense)‘新海16’再生体系基础上,以MSB附加0.1 mg/L 2,4-D及0.1 mg/L KT为基本培养基,采用单因素随机试验研究外植体不同部位、光照及培养基成分等对‘新海16’愈伤组织诱导的影响。结果表明:‘新海16’愈伤诱导的外植体选用下胚轴中上部绿色茎段诱导效果较优;除此之外,愈伤诱导效果最佳的处理为2 000 lx光照强度、90%MSB无机盐成分、3 g/L PVP及1.2~1.4 g/L Gelrite或Phytagel,均能够降低愈伤组织出现白霜现象,对愈伤组织疏松生长有促进作用。本研究通过优化愈伤组织的诱导方法,提高胚性愈伤组织发生率,为‘新海16’高频再生体系提供科学依据。     

2,4-D和6-BA对菊芋愈伤组织诱导的影响

菊芋是一种十分具有开发利用价值的菊科植物,试验以MS培养基为基本培养基,附加不同种类和浓度的植物生长调节物质,对菊芋的块茎进行愈伤组织的诱导培养研究,结果表明:浓度为0.05 mg/L的6-BA的诱导效果最差;6-BA 0.05 mg/L和2,4-D 0.05 mg/L组合有利于愈伤组织的诱导;浓度为0.05 mg/L的2,4-D最有利于愈伤组织的诱导.     

不同因素对贺兰山丁香愈伤组织诱导的影响

利用正交实验设计研究4种因素(6-BA、NAA、2,4-D、外植体)对贺兰山丁香愈伤组织诱导的影响,筛选出最佳的愈伤组织诱导条件为MS 6-BA 0.5 mg/L 2,4-D 0.1 mg/L,其中6-BA的浓度和外植体类型对贺兰山丁香种苗的出愈率有极显著影响(P=0.01).同时发现子叶和胚轴均可作为外植体材料,且以胚轴为外植体在最佳诱导条件下出愈率高达93.3%.     

陇东马牙枣幼胚培养

以陇东马牙枣盛花后50～60d的幼胚为试材,采用L9(3)4正交设计,筛选适宜的幼胚萌发培养基。结果表明,基本培养基、激素配比、蔗糖的含量对幼胚萌发都有显著的影响;其中MS-H培养基的萌发率最高为47.62%;ZT最适宜含量为0.5mg/L,IAA最适宜含量为0.6mg/L;较高的蔗糖含量(50g/L)有利于胚的萌发。幼胚萌发的适宜培养基为MS-H+ZT0.5mg/L+IAA0.6mg/L+LH0.5g/L+蔗糖50g/L+琼脂4.0g/L,剥去种皮,幼胚萌发效果较好。当胚萌发后,将胚从胚乳中或愈伤组织中剥离继续诱导愈伤组织,再经诱导不定芽,生根获得正常植株。