Optimization of Culture Techniques for DH Line in Brassica napus L.

[Objective] The aim of this study is to reveal the disinfectants and disinfection methods,medium components,and embryoid culture method on dissociative microspore culture.[Method] B5 as basic medium appended with different concentrations of sucrose,agar and different hormone combinations was used to optimize the culture technique for DH line in Brassica napus L.[Result] Both the 15 min disinfection of NaClO containing 5% Cl-and 10 min disinfection of 0.1% HgCl2 performed well in disinfection and subsequent embryo production;in the extraction process of dissociative microspores,B5 medium containing 2% sucrose could achieve a good embryo production effect;under dark condition microspores were firstly incubated at 32 ℃ 5-7 d,then at 25 ℃ 12-15 d,and finally transferred to 25 ℃ oscillator(60-65 r/min)for 3-7d,when the embryoid would become full ripeness;1/2MS medium appended with 1.2% agar,0.02% NAA,2.0 mg/L 6-BA,3.4 mg/L AgNO3 and 2% sucrose was helpful for embryoid differentiation and plantlet generation,presenting low degree of browning and slight vitrification.[Conclusion] The results may facilitate DH Line in rape production in large scale and high efficient transformation system.     

甘蓝型油菜DH系培养技术优化(英文)

[Objective] The aim of this study is to reveal the disinfectants and disinfection methods,medium components,and embryoid culture method on dissociative microspore culture.[Method] B5 as basic medium appended with different concentrations of sucrose,agar and different hormone combinations was used to optimize the culture technique for DH line in Brassica napus L.[Result] Both the 15 min disinfection of NaClO containing 5% Cl-and 10 min disinfection of 0.1% HgCl2 performed well in disinfection and subsequent embryo production;in the extraction process of dissociative microspores,B5 medium containing 2% sucrose could achieve a good embryo production effect;under dark condition microspores were firstly incubated at 32 ℃ 5-7 d,then at 25 ℃ 12-15 d,and finally transferred to 25 ℃ oscillator(60-65 r/min)for 3-7d,when the embryoid would become full ripeness;1/2MS medium appended with 1.2% agar,0.02% NAA,2.0 mg/L 6-BA,3.4 mg/L AgNO3 and 2% sucrose was helpful for embryoid differentiation and plantlet generation,presenting low degree of browning and slight vitrification.[Conclusion] The results may facilitate DH Line in rape production in large scale and high efficient transformation system.     

百蕊草无性系建立与瓶外生根研究(英文)

[Objective] The aim of this study is to establish the rapid micro-propagation system in Thesium chinense Turcz.[Method]With stem fragments of wild Thesium chinense Turcz as explants,different culture media were designed to conduct induction culture,strengthening plantlet culture and in vitro rooting.[Result]The optimum medium for inducing clustered shoots was determined to be MS medium appended with 1.5 mg/L 6-BA,0.01 mg/L NAA and 0.3 mg/L 2,4-D;in addition,60 mg/kg ABT was suitable for rooting,by which the percentage of rooted plantlets reached 76.6%.[Conclusion]This study simplified the procedures of tissue culture in Thesium chinense Turcz and enhanced the proliferation rate,providing basis for artificial cultivation and resource protection of Thesium chinense Turcz.     

“胰岛素-转铁蛋白-硒钠”对山羊卵母细胞体外成熟及胚胎发育的影响(英文)

[Objective] The research aimed to enhance culture efficiencies of oocyte and embryo of goat in vitro and to explore serum-free culture system in vitro.[Method] At present,the conventional solutions of oocyte maturation and embryo development in vitro were always added into 1% ITS(Insulin-transferrin-selenium) or using 1% ITS to replace FBS in 2 kinds culture solutions for conducting in vitro cultures of goat oocyte and parthenogenetic embryo.The influences of ITS on their developments were detected.[Result] ITS in maturation liquid of oocytes could not increase oocytes maturation rate but significantly increased blastocyst rate (58.06% vs. 48.19%)of parthenogenetic embryo.If FBS in maturation liquid of oocytes was replaced by ITS, the maturation rate, cleavage rate and blastocyst rate were basically unchanged.Adding ITS into embryo medium could increase blastocyst rate (68.30% vs. 56.82%)of parthenogenetic embryo of goat.If FBS in embryo medium was replaced by ITS,the cleavage rate didn’t change basically,while the blastocyst rate in ITS was obviously lower than that in FBS group(42.33% vs.56.82%).[Conclusion] ITS could promote maturation of oocyte in vitro and early embryonic development, in addition,ITS could replace serum in maturation medium of oocyte as serum-free culture system for conducting relevant researches.     

Somatic Embryogenesis and Plant Regeneration from Leaflets of Fritillaria ussuriensis M.

A method for the production of somatic embryos and subsequent plant regeneration for fritillaria ussuriensis M.is described.Whole leaflet explants,derived from plantlets grown in vitro,formed light yellowith embryogenic calli within one month of culture in the dark.Somatic embryogenesis was obtained after a 28d incubation on MS induction medium supplemented with 2mg/L 2,4-D 0.5mg/L BA,0.5mg/L KT and 500mg/L CH followed by transfer to a second N medium containing 0.5mg/L KT and 100mg/L CH.Somatic embryos were transferred to MS medium with 0.1mg/L NAA placed in the light for regeneration ,After two weeks,mature somatic embryo developed into whole phantlet.     

枸杞花药培养的初步研究(英文)

To investigate the culture technique in anther of Chinese wolfberry,we optimized the culture medium(including hormone combination)and culture conditions.The results showed that calluses were induced from all the six tested Chinese wolfberry materials,but the induction rate of callus varied toward the materials with different genotypes.When the experimental materials were cultured on medium appended with 2,4-D 1.0 mg/L and KT 1.0 mg/L under dark,the callus induction rate reached 20.0 % in this study,and this hormone combination should be the optimum for anther culture of Chinese wolfberry.With MS appended with 6-BA 0.5 mg/L and NAA 0.1 mg/L as differentiation medium and that appended with NAA 0.1 mg/L,the plants could be yielded in 20 days.     

Effects of Different Culture Conditions on Vitrification of Lycium barbarum L. Plantlets in Tissue Culture

The tender stems from new Lycium barbarum L. cultivar "Ningqi 3" released by Ningxia Academy of Agricultural and Forestry Sciences were regarded as explants to investigate the vitrification of Lycium barbarum plantlets in tissue culture under different concentrations of 6-BA,sucrose,agrose,culture temperature,and illumination duration with MS as basic medium. The results show that the conditions for maximal proliferation coefficient and minimal vitrification are as following: the basic medium with 0.2 mg/L 6-BA,3% sucrose and 0.65% agarose; culture at 25 ℃; 12 h/d(daylight lamp,2 000 lx) illumination.     

不同培养条件对枸杞组培苗玻璃化的影响(英文)

The tender stems from new Lycium barbarum L. cultivar "Ningqi 3" released by Ningxia Academy of Agricultural and Forestry Sciences were regarded as explants to investigate the vitrification of Lycium barbarum plantlets in tissue culture under different concentrations of 6-BA,sucrose,agrose,culture temperature,and illumination duration with MS as basic medium. The results show that the conditions for maximal proliferation coefficient and minimal vitrification are as following: the basic medium with 0.2 mg/L 6-BA,3% sucrose and 0.65% agarose; culture at 25 ℃; 12 h/d(daylight lamp,2 000 lx) illumination.     

大豆体细胞胚胎发生系统组织学研究(英文)

Soybean somatic cell could induce the development of embryoid which was similar to embryo morphologically and structurally. Somatic embryogenesis system of soybean was used to conduct genetic transformation of soybean because of its several advantages such as higher transformational efficiency,beetter synchronism and fewer plant chimeras among transgenic plants. After infected with agrobacterium tumefaciens,the initiation,differentiation and development of young cotyledon embryogenic cell of soybean which was cultured on selective culture medium with kanamycin were investigated through histological study. The result showed that somatic embryo was differentiated in non-bud differentiation way. The embryogenic cells were differentiated from epidermis of explant or cells in 1 layer or 2 layers,with the division of embryogenic cells and degradation and disorganization of surrounding cells,the embryogenic cells would form embryoid with analogous suspensor structure.Later,globular embryoid would extrude from epidermis then developed into heart-shape embryo.The experiment was expected to provide theoretical reference for the construction of high transformational system of using plant somatic embryogenesis induced by young cotyledon of soybean.     

Histological Study on Soybean Somatic Embryogenesis

Soybean somatic cell could induce the development of embryoid which was similar to embryo morphologically and structurally. Somatic embryogenesis system of soybean was used to conduct genetic transformation of soybean because of its several advantages such as higher transformational efficiency,beetter synchronism and fewer plant chimeras among transgenic plants. After infected with agrobacterium tumefaciens,the initiation,differentiation and development of young cotyledon embryogenic cell of soybean which was cultured on selective culture medium with kanamycin were investigated through histological study. The result showed that somatic embryo was differentiated in non-bud differentiation way. The embryogenic cells were differentiated from epidermis of explant or cells in 1 layer or 2 layers,with the division of embryogenic cells and degradation and disorganization of surrounding cells,the embryogenic cells would form embryoid with analogous suspensor structure.Later,globular embryoid would extrude from epidermis then developed into heart-shape embryo.The experiment was expected to provide theoretical reference for the construction of high transformational system of using plant somatic embryogenesis induced by young cotyledon of soybean.     

甘蓝型油菜DH系培养技术优化

[目的]研究消毒处理方式、培养基成分和胚状体培养方法对油菜游离小孢子培养的影响。[方法]以B5培养基为基础培养基,添加不同浓度的蔗糖、琼脂及不同激素组合进行试验,对油菜DH系的组培技术进行优化。[结果]含2%Cl-的NaClO消毒15 min与0.1%HgCl2消毒10 min培养效果较好,都能达到良好的消毒和产胚效果;在游离小孢子提取的过程中,B5液体培养基中蔗糖浓度为2%时能达到较好的产胚效果;小孢子培养首先在32℃暗培养5~7 d,然后25℃暗培养12~15 d,最后25℃振荡暗培养3~7 d胚状体就可完全成熟;1/2 MS培养基(附加1.2%琼脂+0.02%NAA+2.0 mg/L6-BA+3.4 mg/LAgNO3+2%蔗糖)有利于胚的分化和苗的形成,其褐化程度小,玻璃化程度低。[结论]该研究结果有助于甘蓝型油菜DH系的规模化生产及高效转化体系的建立。     

2个甘蓝F1小孢子培养中高出胚率的诱导技术研究

以2个优良的F1甘蓝为试材,探讨了高温热激、蔗糖浓度、添加和更换培养液培养、激素种类与浓度配比等因素对提高甘蓝小孢子诱导胚产量的影响.结果表明,2个F1甘蓝材料经24 h 、33℃高温热激处理对小孢子胚胎发生是必要的;小孢子在13%蔗糖浓度培养基(NLN-13)中培养胚产量最高,为1.89胚/蕾;高浓度蔗糖的培养基(NLN-17)培养3 d后添加低浓度蔗糖培养基(NLN-10)培养能大幅度提高胚产量,比一直在13%蔗糖的培养基(NLN-13)培养的胚状体增加255.2%;NLN-13培养基中添加0.1 mg/L 6-BA和0.05 mg/L NAA能够诱导出高胚产量.     

红菜薹游离小孢子培养的影响因素研究

[目的]为蔬菜新品种的选育提供新的途径。[方法]以7个红菜薹品种为试验材料,分别利用6-BA和NAA诱导红菜薹游离小孢子的培养,研究不同诱导剂、游离小孢子密度、活性炭和AgNO3对红菜薹游离小孢子成胚的影响。[结果]6-BA对红菜薹游离小孢子出胚率的诱导效果比NAA明显。分别利用B5培养基和MS培养基对产出的胚进行培养,B5培养基红菜薹生长快,但植株纤细,而MS培养基上的再生植株生长较缓慢。能出胚的培养皿中游离小孢子密度基本上都是5～6个花蕾/皿。红菜薹游离小孢子出胚的培养皿中均添加了活性炭,其适宜添加浓度为0.1g/L。[结论]在培养红菜薹再生植株时,应先用B5培养基,再用MS培养基,同时添加激素进行诱导。     

尾巨桉3个无性系继代增殖培养研究

[目的]研究尾巨桉3个无性系继代增殖培养技术,为尾巨桉苗木快速繁殖提供理论依据。[方法]以尾巨桉3个无性系DH32-26、DH32-29、DH32-28为试验材料,筛选最佳的植物生长调节剂组合、凝固剂、暗培养天数和培养瓶,达到较高的继代增殖系数。[结果]3个无性系继代增殖培养时最适的植物生长调节剂组合为:DH32-26(6-BA 0.5 mg/L+NAA 0.2 mg/L);DH32-28(6-BA 0.4mg/L+NAA 0.2 mg/L);DH32-29(6-BA 0.3 mg/L+NAA0.2 mg/L);DH32-26的继代增殖系数在凝固剂为卡拉胶和琼脂的培养基中差异不大,DH32-28、DH32-29的继代增殖系数在以卡拉胶作为凝固剂的培养基中比在以琼脂作为凝固剂的培养基中有所提高,在生根培养过程中,以卡拉胶作为凝固剂的培养基远比用琼脂作为凝固剂的培养基诱导出根好;继代增殖初期DH32-26、DH32-28需用黑布进行全暗遮光5~10 d,而DH32-29不需全暗培养;用广口瓶继代增殖培养DH32-29可降低苗木生产成本,但DH32-26、DH32-28不适合用广口瓶进行继代增殖培养。[结论]要提高无性系继代增殖系数,需要选取最优的植物生长调节剂组合、凝固剂、暗培养天数和培养瓶。     

不同培养基和泡水时间处理对油茶花粉萌发率的影响

[目的]研究不同处理方式对油茶（Camellia oleifera）花粉萌发率的影响。[方法]油茶品种为望漠油茶和岑溪软枝油茶。共设不同泡水时间和培养基处理。其中,泡水时间分别为0、10、20、30 m in,培养基分别为5%蔗糖＋100 mg/L硼酸＋3%琼脂、10%蔗糖＋100mg/L硼酸＋3%琼脂和20%蔗糖＋100 mg/L硼酸＋3%琼脂。[结果]在不泡水的情况下,望漠油茶花粉萌发最佳培养基是5%蔗糖＋100 mg/L硼酸＋3%琼脂,其花粉的萌发率为80.4%;岑溪软枝油茶花粉萌发的最佳培养基是10%蔗糖＋100 mg/L硼酸＋3%琼脂,其花粉的萌发率为88.5%。在采用相同培养基时,对油茶花粉泡水10~30 m in,花粉的平均萌发率仅为6.3%,最高萌发率为17.9%。[结论]该研究说明泡水对油茶花粉的生命力有极大损害。因此,在油茶人工辅助授粉中不宜采用水粉喷雾法。     

甜(辣)椒花药培养胚状体诱导与植株再生

以10份不同基因型甜(辣)椒为试材进行花药培养,通过对基因型、取蕾时期、低温预处理、热激处理、碳源及外源激素浓度配比等因素的研究,建立有效的甜(辣)椒花药培养胚状体发生体系.结果表明:基因型是限制甜(辣)椒花药培养胚状体诱导的关键因素,不同品种间出胚率差异显著,其中品种003出胚率最高,为10.8%;处于盛花期的花蕾最适于甜(辣)椒花药培养;4℃低温预处理1-3 d有利于胚状体的诱导,以处理2 d的胚状体产率最高;以2%的麦芽糖代替3%的蔗糖能显著提高出胚率和子叶形胚的比率,筛选出适于甜(辣)椒花药培养胚状体诱导的最佳培养基为Ms+0.5 mg/L NAA+1.0 mg/L KT+2%麦芽糖,能有效地提高出胚率并促进植株冉牛;获得了6个基因型的子叶形胚和再生植株.     

柳枝稷的组织培养技术研究

[目的]探索柳枝稷的不同组培条件,优化其诱导和分化培养基。[方法]以柳枝稷幼穗为外植体进行组织培养获得组培苗,建立相应的植株再生系统。[结果]愈伤组织诱导培养基为MS＋5．00mg／L2,4-D＋0．15mg／L6．BA＋3．00％蔗糖＋0．75％琼脂;继代与增殖培养基为MS＋4．00mg／L2,4-D＋3．00％蔗糖＋0．75％琼脂;分化培养基为MS＋0．2mg／L KT＋3．00％蔗糖＋0．75％琼脂;生根培养基为1／2MS＋0．80％蔗糖＋0．70％琼脂。愈伤组织诱导和继代培养阶段培养温度为24℃,在分化和生根培养阶段培养温度为28℃。幼穗外植体的出愈率达95％,愈伤组织增殖率在800％-1000％以上,分化率达80％以上,生根率在98％以上。经炼苗后,获得的组培苗的移栽成活率达95％以上。[结论]采用优化激素搭配的培养基可得到高效的诱导率、分化率和生根率。     

Pollen Viability in Three Xinjiang Hawthorn Species

[目的]探讨适合新疆野生山楂花粉萌发的培养基配方。以及较适的花粉贮藏温度。[方法]在离体培养条件下,采用随机区组设计筛选适合花粉萌发的蔗糖和硼酸浓度;将野生山楂花粉置于不同温度下贮藏6个月,期间采用筛选出的最适培养基对花粉活力进行测定,每10d测定一次。[结果]阿尔泰山楂、红果山楂、准噶尔山楂花粉萌发的较适培养基配方分别为：1％琼脂＋0.05％硼酸＋15％蔗糖,1％琼脂＋0.01％硼酸＋15％蔗糖．1％琼脂＋0.01％硼酸＋20％蔗糖。三者在各自较适培养基上的花粉萌发率分别为30.8％、58.7％、67.2％。且分别极显著高于其它处理。不同贮藏温度条件处理下,阿尔泰山楂、红果山楂、准噶尔山楂花粉在室温条件下分别保存至40、60和70d时完全失去活力;4-5℃条件下．可分别贮藏至90、130和140d时完全失去活力;-18℃条件下．可分别贮藏至130、160和170d时方完全失去活力。[结论]一定浓度的蔗糖、硼酸对野生山楂花粉萌发有促进作用;不同贮藏温度下,三种野生山楂的花粉活力保持时间由长到短均为：-18℃〉4-5℃〉室温。     

辣木快速繁殖体系

研究通过对辣木种子消毒方法、增殖阶段6-苄氨基腺嘌呤(6-BA)浓度、生根阶段吲哚丁酸(IBA)浓度、移栽炼苗时间进行筛选,确定辣木种子适宜的消毒方法为75%的酒精浸泡30s后用1%NaClO消毒15 min;增殖培养基为MS+1.0 mg/L 6-BA+30g/L蔗糖+7.0g/L琼脂(pH5.8);生根培养基为1/2 MS+0.4mg/L IBA+30g/L蔗糖+7.0g/L琼脂(pH5.8);移栽时炼苗6d较适宜。由此建立了辣木快速繁殖体系,为规模化生产辣木提供参考。