胡萝卜与柴胡的体细胞杂交及其再生愈伤组织的同工酶鉴别

目的:用体细胞杂交技术将中药柴胡的药效成分基因转入胡萝卜,为中药材的种质资源开发、优化和良种选育提供理论与实验数据。方法:取狭叶柴胡的原生质体经300μW·(cm2)-1照射O,1,2min后分别与胡萝卜原生质体用PEG(聚乙二醇)法进行体细胞杂交,对获得的再生愈伤组织进行形态学、同工酶分析。结果:初步确正9个克隆为狭叶柴胡与胡萝卜的体细胞杂种。结论:为进一步分析杂种中柴胡药效成分的含量及筛选出高药效成分含量的细胞系奠定基础。

甜菜原生质体培养再生愈伤组织

国外曾报道利用悬浮培养细胞分离、培养甜菜原生质体,结果分别得到多细胞团或愈伤组织。国内有关甜菜原生质体培养尚未见报道。本文报道对悬浮细胞来源的甜菜原生质体的培养结果。

不同干燥时长对小麦成熟胚愈伤组织再生进程的影响

小麦组织培养和细胞培养对许多领域的研究具有至关重要的作用,尤其是在突变体的筛选和植株移植方面有着举足轻重的作用。植株再生是小麦组织培养中的关键环节,它已经成为离体筛选有益突变体和转基因研究的基础。为了提高小麦愈伤组织再生的能力,本研究利用开麦18、开麦20、开麦22和开麦1502等4个小麦品种成熟胚愈伤组织为试验材料,在一定的光照和温度条件下经过15周的继代培养后,分别采取6 h、12 h、24 h、48 h、72 h干燥处理,探索其干燥率、吸胀率和再生率。结果表明,在分化前对小麦成熟胚伤组织进行吸湿干燥处理,可以普遍提高愈伤组织的再生率,加快再生进程;4个品种的再生率均在干燥48 h时最高,以开麦1502诱导效果最好,其再生率达21.63%。

小麦成熟胚愈伤组织诱导及再生影响因素初探

以2个小麦品种成熟胚为外植体进行离体培养,研究了不同取胚方式、诱导培养基中不同2,4-D浓度、分化培养基中不同2,4-D浓度与KT浓度组合对愈伤组织的诱导率、直接成苗/根率及再生成苗率的影响。结果表明:刮胚法的再生苗率最高、直接成苗/根率最低;在2个基因型条件下,2,4-D浓度在2 mg·L-1条件下,愈伤组织出愈率均最高;在分化培养基中,MS+2,4-D 2 mg·L-1+KT 2 mg·L-1其再生率最高。

根癌农杆菌介导转化川草二号老芒麦胚性愈伤组织

以川草二号老芒麦成熟种子为外植体,经过对培养基的筛选和培养条件优化,建立了愈伤组织再生系统。转化载体为pCAMBIA1304质粒,其T-DNA上携有潮霉素抗性基因(hptII)和类产碱假单胞菌杀虫蛋白基因(ppIP),经根癌农杆菌EHA105介导转化结构致密、颗粒状、黄白色的胚性愈伤组织。通过潮霉素筛选和对抗性植株进行分子检测,获得了转基因植株。同时优化了农杆菌遗传基因转化的参数,建立了农杆菌介导的川草二号老芒麦程序化转基因方案。

药用植物莪术的组织培养快速繁殖与植株再生的研究

通过茎尖培养实现了中药莪术试管苗的快速繁殖并从试管苗基部诱导出了愈伤组织。实验表明 :以 MS培养基为基本培养基 ,附加 ZT 4.0 mg/ L + IAA 0 .5～ 1.0 m g/ L适于丛生芽的诱导与增殖 ;附加 2 ,4- D 1.0 mg/ L+ KT4.0～ 6 .0 m g/ L 适于愈伤组织的诱导 ;附加 2 ,4- D1.0～ 2 .0 mg/ L + KT0 .2～ 0 .5 mg/ L 适于愈伤组织的继代培养 ;附加 KT 4.0 m g/ L + NAA 0 .2 m g/ L适于愈伤组织的分化。

双亚5号亚麻与罗布麻原生质体解离及体细胞杂交

【目的】探索双亚5号亚麻与罗布麻原生质体制备、电融合和培养的适宜条件,为进一步深入进行亚麻与罗布麻体细胞杂交育种研究奠定基础。【方法】以已建立的双亚5号亚麻和罗布麻胚性细胞悬浮系为材料,制备原生质体并进行电融合,培养。【结果】制备亚麻和罗布麻原生质体的最佳渗透剂分别是5.6%蔗糖和0.7mol/L甘露醇;游离亚麻和罗布麻原生质体的最适酶组合都是0.5%CellulaseOnozukaR-10+1.0%CellulaseOnozukaRS+0.2%PectolyaseY-23;最佳酶解时间分别是4和3h;亚麻细胞最佳继代时间是转瓶后的第3d,在此条件下原生质体存活率均为92.0%,产量分别达到1.24×106和22.4×106个/mL;2种原生质体电融合的最佳参数依次为:AC8v/cm,持续25s—DC1500v/cm,脉冲宽度为60μs,脉冲次数1次—交流电时间为9s,重复次数2次。在此条件下融合率达到20.5%。【结论】亚麻与罗布麻原生质体在一定的条件下可实现体细胞杂交。

Establishment of Rapid Propagation Technique for Cotyledon of Solanum torvum

[Objective] To research the mass propagation system for cotyledon of Solanum torvum. [Methods] With cotyledon of S. torvum as the research object, ef- fects of hormone combination on callus induction and adventitious buds differentia- tion of S. torvum were researched. [Results] With cotyledon of S. torvum as the ex- plants, the optimal culture medium for callus induction and adventitious buds differ- entiation was MS＋2.0 mg/L 6-BA＋0.3 mg/L NAA. The induction rates of callus and adventitious bud reached 100% and 85%, respectively. The number of average buds was 6. The optimal culture medium for the induction of adventitious roots was MS＋ 0.3 mg/L IAA. The rooting rate reached 100% and the number of average roots was 9. [Conclusions] One-step induction of callus and adventitious buds simplified the mass propagation system, and enhanced the test test efficiency.s

不同白桦优树组培快繁体系的筛选

用已建立的白桦离体再生培养体系对20个白桦杂交优树(Betula platyhylly×Betula pendula)腋芽进行了离体培养,比较它们腋芽再生和愈伤组织再生情况,并分析了无性繁殖的变异情况。结果表明:20个优树腋芽启动率存在的差异较大,最高的为4号和7号优树,为75.0%,最低的为17号优树,仅24.4%;3号和16号优树的启动率较高,分别为63.1%、61.3%;3号和16号优树在腋芽增殖和愈伤组织再生途径中长势都比其它植株好,选择它们用于工厂化育苗,可以满足造林和生产需要。

Callus induction from leaves of different paulownia species and its plantlet regeneration

The experiment was carried out on five different species of Paulownia for callus induction from leaves. MS medium was adopted as basic medium, and from different combinations of NAA and BA the suitable media were determined for callus induction, bud differentiation, and root differentiation of five different species. MS+0.5NAA+4BA, MS+0.3NAA+2BA, MS+0.5NAA+4BA, MS+0.3NAA+6BA, and MS+0.3NAA+8BA were suitable media of callus inductions of leaves, respectively, for Paulownia tomentosa, Paulownia australis, Paulownia fortunei, Paulownia elongata and P. tmentosa x P. fortunei, and MS+0.3NAA+12BA, MS+0.3NAA+12BA, MS+0.5NAA+12BA, MS+0.5NAA+12BA, and MS+0.7NAA+12BA were suitable media for bud differentiation from leaf callus respectively for above five species. The rooting media was determined as 2MS+0.1NAA, 1/2MS+0.1NAA, 1/2MS, 1/2MS+0.3NAA, and 1/2MS+0.5NAA. These results provide reference data for breeding new fine va-rieties with different kinds of Paulownia protoplasts fusions.

Discussion on the Regeneration Technology of Gazania rigens L.Leaves

[Objective]The research aimed to study the regeneration technology of Cazania rigens L.leaves and screen out the optimum medium formula for the regeneration of Cazania rigens L.leaves.[Method]Using Japan imported C.rigens leaves as materials,the orthogonal test was made for the callus and adventitious buds induction in MS medium with different kinds and concentrations of hormones.The optimum medium formula for the regeneration of C.rigens leaves were screened out.[Result]On the medium of MS ＋ 0.8-1.0 mg/L TDZ ＋ 0.05-1.0 mg/L NAA,compact type and bright green calli were formed.When the leaves were inoculated on the medium of MS ＋ 0.5-1.0 mg/L TDZ ＋ 0.05-1.0 mg/L NAA,many adventitious shoots can be induced and the induction rate reached 100%.When strong adventitious shoots with the height of 2.0-3.0 cm were transplanted into the medium of 1/2 MS ＋0.1 mg/L NAA,the rooting situations were good and the rooting rate was 100%.[Conclusion]The research provided a new way for the rapid propagation of C.rigens and laid the foundation for the genetic transformation and new varieties breeding of C.rigens.

A Genetic Transformation System for Rosa multiflora Thunb. var. cathayensis Rehd. et Wils through Callus Induction

An efficient genetic transformation system is a preparation for Rosa multi-flora Thunb. var. cathayensis Rehd. et Wils to diversify its flower color through ge-netic engineering. We firstly optimized the explants and culture conditions on callus induction, hormone concentrations and dark period of culture time on bud differentia-tions in particular, with sterilized seedlings to establish the regeneration system of R. multiflora. It showed that callus induction frequency reached 100% after the ex-plants being cultured in dark for 21 d when MS was chosen to be the initial culture medium. The bud differentiation rate was 48% after cal i being cultured under dark for 8 d on MS medium supplemented with TDZ （1.5 mg/L） and NAA （0.05 mg/L）. The cal i was used as the explants that were infected with Agrobacterium tumefa-ciens harboring a DFR-RNAi construct. The transformation rate reached as high as 50%. The establishment of a highly efficient rose gene transformation system out-lined in this report is prerequisite for genetic improvement in rose flower colors.

Direct Regeneration of Inflorescence from Callus in Dracaena fragrans cv. Massangeana Hort.

以花叶千年木 (Dracaenafragranscv .MassangeanaHort.)的花被筒、花序分枝轴和花序轴为外植体成功地诱导了花序的直接再生。3种外植体首先在MS附加 1.0mg L 6_BA和 0 .5_0 .8mg L 2 ,4_D的培养基上诱导形成愈伤组织 ,然后转移到MS附加 0 .5mg L 6_BA和 0 .0 0 5～ 0 .5mg L 2 ,4_D的培养基上分别诱导了花序的直接再生。

优良白桦单株离体繁殖性状的筛选

用已建立的白桦离体再生培养体系，对10个优良白桦母树单株的腋芽进行离体培养，比较其腋芽再生和愈伤组织再生情况，分析单株间无性繁殖的变异情况。结果表明：10个无性系的腋芽启动率都较低，差异不是很大，均在50％～60％，其中m-8和m-1启动率较低，但在腋芽增殖和植株再生途径中，长势都较其它无性系好，所以选择m-8和m-1号无性系用于工厂化育苗、造林和生产需要。

长白山濒危树种刺楸组培快繁技术研究

以长白山刺楸人工林选择的刺楸优良个体休眠枝条为试验材料,进行刺楸组培快繁各个阶段最适培养基的筛选。结果表明,刺楸组织培养的最适外置体为叶柄;愈伤组织诱导的最适培养基为WPM+2,4-D0.50mg/L+NAA0.1 mg/L,诱导率96%;愈伤组织诱导再分化最适培养基为WPM+KT2.0 mg/L+6-BA0.50 mg/L+NAA 0.1 mg/L,分化率达82%;不定芽继代培养最适培养基为WPM+KT1.0 mg/L+6-BA0.50 mg/L+NAA0.1 mg/L,增殖系数为3.69;不定芽生根培养最适培养基为WPM+NAA 0.5 mg/L,生根率达到85%。

Effects of 2,4-D and 6-BA on Callus Induction and Plantlet Regeneration from Mature Embryos of Hsien Rice

[ Objective] In order to study the effects of 2,4-D and 6-BA on callus cultivation from mature embryos of hsien rice. [ Method] 2,4-D and 6-BA were set at different concentrations in callus induction and differentiation mediums to study their effects on callus induction, seedling formation and regenerated seedlings rooting. [ Result] In the callus induction medium treated with 0.5 mg/L 2,4-D, the callus induction effects on the varieties like Jiayu 948, Yanghui 559, Yangxian 6547, Zhong＇erruanzhan, Minghui 86, Guanghui 998 and Zunxian 3 were the best; If 0.2 mg/L 6-BA was added into the callus induction medium containing the optimum level of 2,4-D, there was no obvious effect on induction rate of callus, but the differentiation and seedling of callus were inhibited; If the concentration of 6-BA was reduced appropriately in the differentiation medium, the seedling rate of callus would be not only no decreased but increased, meanwhile the quality of regenerated plants would be improved. [ Conclusion] The study results provided some references for the reasonable uses of 2,4-D and 6-BA in callus culture of hsien rice.

Optimization of Regeneration System of Leaf and Stem of Populus euramericana 108

[Objective] The aim was to optimize the culture conditions for asexual reproduction system of Populus euramericana 108.[Method] Orthogonal designs were adopted optimize the culture conditions of the regeneration system for direct differentiation from leaves and induced callus from stems of P.euramericana 108 aseptic seeding.[Result] Leaves of P.euramericana 108 directly regenerated and differentiated under illumination,while stem segments preferred to regenerate and differentiate through callus induction under illumination.The differentiation medium of adventitious buds from leaves was MS medium （agar 7.0 g/L,pH 6.0,sucrose 20 g/L） added with 0.6 mg/L 6-BA and 0.2 mg/L NAA; callus induction medium of stem segments was WPM solid medium added with 0.75 mg/L KT and 1.5 mg/L 2,4-D.Rooting induction medium for adventitious buds was WPM solid culture （sucrose 30 g/L） added with 2.0 mg/L IBA.[Conclusion] The culture conditions for regeneration system of differentiation from leaves and induced callus of stems were optimized,which provides basis for the construction of tissue culture and genetic transformation system.

Optimization of plant regeneration system in vitro culture in wheat

Studies were carried out to establish an efficient regeneration system of three bread wheat cultivars. Results showed induction medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) had a higher plantlet regeneration frequency than Piclorm, with an average frequency of 54% in all treatments. Optimal condition for different genotypic rice was as following: induction medium (MSS 3AA/2) with 0.5 mg L-1 2,4-D, regeneration medium (R) with 0.01 mg L-1 2,4-D and 3 mg L-1 KT. The average regeneration frequency reached 83.3% under the condition. Correlation analysis showed that root differentiation, in different level, correlated with green spot regeneration, and with the number of regenerated plants per callus. No correlation was found between green spots regenerated and the numbers of plants regenerated per callus.

Regeneration of Leaves in vitro in Cherry Rootstock Colt

Plants were regenerated from leaves of cherry rootstock Colt by two methods,the frequencies were 48 3% and 21 3%.Leaves were cultured on MS medium supplemented with NAA 1 0mg/L,KT 3 0mg/L and ZT 0 25mg/L.After 5～7 days leaves dedifferentiated,and formed callus.About 25 days later,callus grew into greenish or pink compact ones,the induction frequency was 100%.The color and structural feature of callus depended on the medium,culture condition and physiology phase of leaves.To induce callus,NAA was the main factor.Leaves rooted in medium only with NAA.In order to inhibit rooting and make callus grow fast,KT and ZT were added to the medium.Leaves cultured without light could quickly form callus during the initial stage of dedifferentiation.Young leaves dedifferentiated more easily than old leaves.After had been cultured about 50 days,callus were transferred to MS medium supplemented with NAA 0 2mg/L,IAA 0 5mg/L,6 BA 0 5mg/L,KT 1 0 mg/L and GA 0 5mg/L,and redifferentiated at a frequency of 21 3%.The callus of Colt leaves were difficult to redifferentiate.GA was important in the redifferentiation of callus.The result showed that callus could redifferentiate in all the mediums with GA,and could not redifferentiate without GA.The ability of redifferentiation of big callus was higher than that of small ones.And the physiology character of callus was also important.High regeneration frequency was obtained from greenish or pink compact callus.The frequency of regeneration of petioles was higher than that of leaves.Leaf petioles' regeneration need high concentration of cytokinin(more than 5 0mg/L).Leaf petioles were cultured on MS medium supplement with 6 BA 6 0mg/L,NAA 1 0mg/L and GA 0 5mg/L,and regenerated at a frequency of 48 3%.