不同基因型对小麦成熟胚愈伤组织诱导及植株再生的影响

以26种不同基因型小麦成熟胚为外植体，采用1种诱导培养基、3种分化培养基，研究了不同基因型间愈伤组织诱导及植株再生的遗传差异．结果表明，不同基因型间出愈率及植株再生率差异显著，豫麦21号在26个小麦品种中表现最佳，3种分化培养基的分化效果差异明显．     

小麦成熟胚愈伤组织诱导及分化的研究

以小麦成熟胚为外植体，在ＭＳ＋２．４－Ｄ１．０ｍｇ／Ｌ＋６－ＢＡ１．０ｍｇ／Ｌ培养基上，愈伤组织诱导率为８３．１％；不同品种间诱导率有较大的差异，但愈伤组织的质量与诱导率的高低无关，在ＭＳ＋ＫＴ１．０ｍｇ／Ｌ＋ＩＡＡ０．１ｍｇ／Ｌ培养基上，分化率可达４４．４％。     

小麦成熟胚愈伤组织的诱导及植株再生体系的研究

以小麦的成熟胚为外植体,通过胚性愈伤组织诱导进行植株再生.研究结果表明,在MS+2,4-D(3.0 mg/L)+VB1(1.0 mg/L)的培养基中,成熟胚可较高频率的诱导产生愈伤组织,诱导率为72.3%,但愈伤组织质量较差;在低浓度2,4-D和添加了ABA的继代培养基中,愈伤组织质量明显改善,胚性愈伤组织增加.筛选继代的胚性愈伤组织置于分化培养基MS+6-BA(1.0mg/L)+KT(4.0 mg/L)+NAA(0.3 mg/L)中,分化率达73.18%.该研究建立了一套有效的以成熟胚为外植体的小麦组织培养再生体系.     

小麦胚愈伤组织的诱导和植株再生

对3个小麦品种的成熟胚及幼胚进行培养，研究不同基因型、培养基对愈伤组织诱导及分化的影响。结果表明，不同培养基对小麦愈伤组织诱导有一定影响；不同基因型的小麦愈伤组织诱导、分化及生根的能力有一定差别。陕280愈伤组织诱导率及分化率最高。同一品种不同外植体经诱导后愈伤组织产生的能力有所不同，同一品种愈伤组织的诱导率幼胚比成熟胚愈伤组织的高。     

小麦成熟胚愈伤组织的诱导和植株再生

从完整的小麦种子和它的离体胚成功地诱导出了愈伤组织。愈伤组织诱导和继代培养基中2,4-D、VB_1和肌醇的浓度影响植株再生频率。在分化培养基中加入500mg/L 水解酪蛋白促植再生。KT 对于愈伤组织的生长和分化有抑制作用。不同品种愈伤组织的植株再生能力相差很大,京红8号的植株再生频率较高,为76%。     

不同预处理对小麦成熟胚愈伤组织形成的影响

研究了低温、CaCl2 、PEG 3种预处理对 18个小麦品种成熟胚愈伤组织形成的影响 ,结果表明 :不同预处理对小麦成熟胚愈伤组织的形成有显著影响。PEG处理对小麦成熟胚愈伤组织的形成具有显著促进作用 ;低温和CaCl2 溶液处理对小麦成熟胚愈伤组织的影响和小麦品种关系密切 ,品种间差别较大。     

小麦成熟胚愈伤组织的诱导及分化

以4个基因型不同的小麦品种成熟胚为外植体进行离体培养,研究不同质量浓度2,4-D对愈伤组织诱导的影响;以陕253为外植体研究不同KT与NAA配比对小麦成熟胚愈伤组织分化的影响.愈伤组织培养基中2,4-D质量浓度为2～4 mg/L时,小麦成熟胚愈伤组织出愈率最高,品质最好.小麦对水分敏感程度与成熟胚愈伤组织的出愈率及品质...     

小麦成熟胚愈伤组织的诱导与分化

[目的]筛选再生能力强的基因型小麦品种,建立高效的小麦成熟胚高频植株再生系统。[方法]以小麦品种抗冻早-30、百麦4411和06-4046的成熟胚为外植体,以MS为基本培养基,添加不同激素,组配诱导、继代、分化和生根培养基,通过胚性愈伤组织诱导。探讨影响小麦愈伤组织诱导和分化的因素。[结果]2,4-D浓度为1—5mg／L时对3个小麦品种愈伤组织的诱导差异不大,诱导率均高于80％,但2,4-D浓度为2mg／L时更有利于诱导胚性愈伤组织,诱导率在90％以上;在抗冻早-30和百麦4411分化培养基中加入2mg／L KT有助于提高分化率和再生率,在06-4046分化培养基中加入0．5mg／L KT对小麦成熟胚的分化和再生苗比较有利。[结论]在MS培养基中添加2mg／L2,4-D和一定量的KT可促进小麦成熟胚愈伤组织的诱导和分化再生苗。     

小麦成熟胚愈伤组织诱导研究进展

小麦品种的遗传改良一直倍受重视,利用小麦成熟胚为外植体进行愈伤组织培养越来越受到人们的关注。本文主要对影响小麦成熟胚愈伤组织诱导的诸多因素进行了讨论。     

玉米自交系成熟胚愈伤组织诱导和再生体系研究

[目的]研究玉米成熟胚组织培养条件和植株再生频率的影响因素,建立玉米成熟胚高效再生体系.[方法]以玉米成熟胚为材料,通过相关影响因子对成熟胚外殖体愈伤组织诱导、继代及分化培养影响的研究.[结果]2,4-D浓度在1.0-2.0 mg/L,NAA浓度为0.5-0.6 mg/L明显促进成熟胚愈伤组织生长;6-BA浓度为0.15mg/L时适合愈伤组织的继代培养;新自523在三个自交系中愈伤生长质量较好,并且容易产生胚性愈伤,早21愈伤组织较少,在0.5 mg/L NAA、1.5 mg/L6-BA、100 mg/L AgNO3浓度下早21分化能力和新自523相当,且两者都容易生根,再生苗频率达到30;-40;.B73分化和生根能力较弱.[结论]利用玉米成熟胚诱导产生愈伤、分化并产生再生植株是继幼胚再生体系有利的补充.     

Dicamba and Sugar Effects on Callus Induction and Plant Regeneration from Mature Embryo Culture of Wheat

To establish a highly efficient plant regeneration system for wheat genetic transformation, the effects of three different concentrations of dicamba and two different sugar types on callus induction and plant regeneration from mature embryo cultures were evaluated. Callus induction and plant regeneration were obtained from mature embryos of two commercial cultivars Zhoumai 18 and Yumai 34 （Triticum aestivum L.） cultured on L3 basal medium. The results showed that the efficiency of mature embryo culture was significantly influenced by the genotypes, sugar types and dicamba concentrations. 4 mg L^-1 dicamba proved the best effective for inducing embryogenic callus and also gave the highest proportion of plants regenerated across the two cultivars. Substitution of maltose by sucrose significantly improved the plant regeneration efficiency in both cultivars. There was a significant interaction between genotype-by-sugar types, and sugar types-bydicamba concentrations. Overall, Zhoumai 18 gave the highest frequency of plant regeneration （82.65%） when dicamba concentration was 4.0 mg L^-1 and with sucrose in initial callus induction. These results will facilitate genetic transformation work with elite wheat.     

Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture

To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm （endosperm-supported culture, ES）, the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm （non-endosperm-supported culture, NES）. This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.     

Effects of Environmental Temperature on the Regeneration Frequency of the Immature Embryos of Wheat （Trificum aestivum L.）

The immature embryos （IEs） of wheat are the most widely used tissues for in vitro culture and genetic transformation due to its high regeneration competency. However, this explant can only be maintained in 4℃ daily cooler for a short period time for its use in plant tissue culture or transformation experiments. This study aimed to investigate the effects of environmental temperature, cryopreservation storage temperature, and heat shock culture （HSC） temperature on the regeneration frequency of wheat IEs. Results indicated that environmental temperature significantly affected the induction of embryonic calli. The optimum total accumulated temperature （TAT） during the time of anthesis and sampling for regeneration of these tissues was around 280℃ for spring wheat type cv. CB037 and approximately 300℃ for winter wheat type cv. Kenong 199. Regeneration ability obviously declined when the highest environmental temperature was over 35℃ for 1 d or a high temperature between 30 and 33℃ lasted for 5 d during anthesis and sampling. This finding was verified by culturing the freshly isolated IEs under different temperatures from 29 to 37℃ in different controlled growth incubators for 5 d; the IEs almost completely lost regeneration ability when the temperature rose to 37℃. Cryopreservation of-20℃ caused the wheat samples lost ability of producing callus or embryonic callus in a few days, and cryopreservation of-10℃ more than 10 d made the regeneration potential of the tissues dramatically declined. Comparatively, the temperature that best maintained high regeneration ability was -5℃, at which the materials can be maintained for around 1 mon. In addition, the preservation of the immature samples at -5 or -10℃ inhibited the direct germination of the IEs, avoiding the embryo axis removing process. Our results are useful for ensuring that field collection and cryopreservation of the wheat IEs are done correctly to enable tissue culture and genetic transformation.     

小麦成熟胚离体培养研究

以美国小麦品种Overley成熟胚作为外植体进行离体培养,研究外源激素对小麦成熟胚诱导愈伤组织、分化出苗以及试管苗增殖的影响。试验结果表明,在MS培养基中添加2,4-D3mg·L-1愈伤组织诱导频率较高,成愈率为84.0%;在添加有KT2～3mg·L-1的MS分化培养基中愈伤组织分化率较高,可达85%以上;将试管苗转接至添加0.2mg·L-1的NAA和0.5～2.0mg·L-1的6-BA的培养基中,试管苗有一定的分化增殖,但最高分化率仅为33.3%,增殖系数2.05。     

十二个小麦新品种成熟胚再生性能与农杆菌侵染敏感性评价

小麦成熟胚培养再生率比较低,尤其对优良小麦品种成熟胚再生性和农杆菌侵染敏感性还缺乏研究,一定程度阻碍了转基因小麦有效开展和产业化进程。以扬麦18、扬麦15、扬麦16、新冬18、小偃166和周麦22等12个优良小麦新品种为材料,研究了其成熟胚再生率及其对农杆菌侵染的敏感性。结果表明,12个小麦新品种成熟胚再生率0~35.3%,周麦27最高,扬麦19次之（27.9%）,新冬18、石麦B07-4056分别为211%和20.0%,其余品种再生率均低于20.0%;12个小麦品种成熟胚愈伤组织农杆菌侵染后GUS基因瞬间表达率0~33.3%,周麦18最高,扬麦15次之（23.8%）,扬麦18、周麦22、周麦27和石麦B07-4056虽有表达但频率较低（6.7%~9.1%）,其余品种没有观察到GUS基因瞬间表达。认为周麦27、扬麦19等品种适合进行成熟胚培养,周麦18、扬麦15等品种成熟胚适合进行农杆菌转化。优良小麦品种成熟胚再生性能和农杆菌敏感性评价为进一步利用细胞工程和基因工程途径改良小麦奠定了基础。     

水稻成熟胚愈伤组织诱导及其植株再生

利用水稻成熟胚为外植体,接种在NB培养基上,分别附加不同的外源激素,以诱导愈伤组织并促其分化,最终获得水稻再生植株.结果表明:2,4-D对愈伤组织诱导起决定性作用,当它与低浓度的6-BA配比时更有利于前期愈伤组织的诱导,而高浓度的6-BA则有利于后期芽丛的分化.     

不同培养基对小麦幼胚再生能力的影响

以安徽省小麦主栽品种扬麦87158、安农98005、安农92484三个品种的小麦幼胚为试验材料,接种于SD2和MM2种培养基上,探讨不同培养基对不同基因型小麦幼胚再生能力的影响。通过对其愈伤组织诱导率、愈伤组织的质量和绿苗分化率的比较,初步证明SD2培养基对小麦幼胚的再生能力效果好,可以作为转基因小麦育种中的培养手段。     

Establishment and Optimization of the Regeneration System of Mature Embryos of Maize (Zea mays L.)

A reliable system was developed for regeneration from mature embryos derived from callus of four maize inbred lines (Liao 7980,Dan 9818,Dan 340,and Dan 5026).The protocol was mainly based on a series of experiments involving the composition of culture medium.We found that 9 μM 2,4-dichlorophenoxyacetic acid in MS medium was optimum for the induction of callus.The induction frequency of primary calli was over 85% for four inbred lines tested.The addition of L-proline (12 mM) in subculture medium significantly promoted the formation of embryogenic callus but it did not significantly enhance growth rate of callus.Efficient shoot regeneration was obtained on regeneration medium containing 2.22 μM 6-benzylaminopurine in combinations with 4.64 μM Kinetin.Regenerated shoots were rooted on half-strength MS medium containing 2.85 μM indole-3-butyric acid.This plant regeneration system provides a foundation for genetic transformation of maize.     

草坪型高羊茅成熟种子胚性愈伤组织诱导及植株再生

以5种草坪型高羊茅成熟种子为外植体材料,探讨外植体消毒和不同激素组合对愈伤组织诱导、继代培养以及植株分化的影响。结果表明,不去种皮与去种皮相比,能显著提高高羊茅成熟种子发芽率和出愈率。猎狗5号和野马2号在添加2,4 D10.0mg/L的培养基中愈伤组织诱导率最高;新秀和颂歌的出愈率在2,4 D浓度为8.0mg/L时达到最高;野火的出愈率在2,4 D浓度为4 0～10 0mg/L时变化不大,仅为10.5%～11.2%。用3.0g/L的水晶洋菜(Gelrite)代替琼脂做培养基疑固剂可促进愈伤组织生长及向胚性愈伤组织转化;愈伤组织在含BA2.0mg/L和NAA0.5mg/L的MS培养基上植株分化率较高,其中猎狗5号在此培养基上的愈伤组织植株分化率达72.8%。