无核葡萄胚挽救培养污染防治的初步研究

无核葡萄是当今国际葡萄消费和生产的重要方向之一,胚挽救技术是无核葡萄育种的重要手段;无核葡萄胚珠离体培养中,污染现象时有发生,严重阻碍了无核葡萄胚挽救育种的效率。以"皇家秋天"、"红宝石无核"、"无核翠宝"等种子败育型无核葡萄品种自然授粉幼果为材料,进行的无核葡萄胚挽救培养过程中胚珠内胚培养阶段污染影响因素及抑菌培养试验结果表明,75%乙醇浸泡30s+1%HgCl2溶液浸泡6min的消毒处理对无核葡萄胚珠内培养的效果较好;胚珠培养的接种密度对污染有一定的影响;胚形成培养基MM4+香蕉泥500mg·L-1添加0.1g·L-1高锰酸钾的抑菌效果较好。     

淡黄花百合与东方百合''如意''远缘杂交的研究

以淡黄花百合（父本）和东方百合‘如意’（母本）为亲本，采用离体受精的方法和胚挽救技术进行了两种百合的远缘杂交试验研究。结果表明，采用切割花柱的方法，获得了较高的子房膨大率；在胚珠培养过程中，用甘露醇和山梨醇代替部分蔗糖并加入0．5mg／mL6-BA和0．1mg／mLNAA的培养基有利于胚的发育，其有胚率为1％，并成功获得了杂种植株。     

淡黄花百合与东方百合‘如意’远缘杂交的研究

以淡黄花百合（父本）和东方百合‘如意’（母本）为亲本,采用离体受精的方法和胚挽救技术进行了两种百合的远缘杂交试验研究。结果表明,采用切割花柱的方法,获得了较高的子房膨大率;在胚珠培养过程中,用甘露醇和山梨醇代替部分蔗糖并加入0．5mg／mL6-BA和0．1mg／mLNAA的培养基有利于胚的发育,其有胚率为1％,并成功获得了杂种植株。     

设施栽培无核葡萄胚珠、胚发育动态

对设施栽培不同熟期的无核葡萄品种(品系)及其杂交组合进行果实、胚珠、胚的生长发育动态进行研究,得出不同品种(品系)胚珠、胚败育时期和胚珠数量的变化规律,确定胚珠、胚发育的最佳时期,为胚挽救的最佳接种时期提供依据。     

饲料型四倍体刺槐幼胚离体培养挽救技术研究

遗传变异丰富、具有重要遗传育种价值的饲料型四倍体刺槐幼胚自然败育率很高,研究其离体培养挽救技术具有重要的遗传育种学意义。以处于不同发育时期的饲料型四倍体刺槐幼胚为材料,进行了其离体培养促萌试验,试验结果表明:胚珠的适宜离体培养时期为花后30 d,萌动率47.7%;胚珠的适宜萌动培养基为Nitsch 6-BA 0.5 mg/L IBA 2 mg/L GA30.5 mg/L;筛选出的最佳愈伤组织诱导培养基上得到的愈伤组织诱导率为38.6%,并初步诱导出了不定芽;愈伤组织切片表明,饲料型四倍体刺槐幼胚愈伤组织的增殖生长以内起源为主。     

科间远缘杂交种-银白杨×白榆

1978年,辽宁省杨树研究所开始进行银白杨×白榆科间远缘杂交试验研究。利用蒙导技术克服远缘杂交障碍、子房离体培养挽救败育幼胚,成功获得了银×榆F1的完整植株。经形态、亲本及子代AFLP标记测试得出,银×榆属于偏母本的融合型杂种,且银×榆具有抗旱、抗虫等优良特性,是一个比较特殊的珍稀物种资源。     

桂花离体胚的组织培养

为给桂花的良种快繁奠定基础,采用不同培养基和激素对金桂胚的组织培养进行了初步研究。在胚苗培养中,进行了胚的初代萌发诱导培养、胚苗的伸长培养和生根培养,其最佳培养基配方分别为:MS (1.00~2.00)mg.L-16-BA (0.00~0.05)mg.L-1NAA;B5 2.00 mg.L-1GA;1/2MS 3.00 mg.L-1NAA。还进行了胚的愈伤组织初代诱导培养和胚苗茎段初代萌发诱导培养,其最佳培养基配方分别为:LMC (0.12~0.30)mg.L-12,4-D;B5 1.00 mg.L-16-BA 0.05 mg.L-1NAA。     

I-69杨与小叶杨杂交种胚培养研究

本研究利用组织培养技术对进行培养挽救,以为了提高育种成功率.试验结果表明:外植体的最佳消毒方法为75％酒精消毒30 s后在0.1％升汞处理8min,幼胚的成活率为90％.随着外植体消毒时间增加,幼胚成活率先上升后下降.幼胚最佳萌发培养基为1/2MS+ BA0.1 mg·L-1+ NAA0.01 mg·L-1.最佳继代增殖培养基为MS+ BA0.1 mg· L-1+ NAA0.01 mg·L-1,增殖系数达到6.33.对不定芽增殖最有利条件为:pH值6.0、琼脂浓度6g·L-1和蔗糖浓度30 g·L-1.最佳生根培养基为1/2MS+ IBA0.2 mg·L-1,生根率为93.2％.     

落叶松体细胞胚发生研究进展

以体细胞胚发生的发展阶段为线索，对落叶松胚性培养物的诱导、胚性愈伤组织的保持和增殖、体胚成熟、体胚的萌发进行了全面的论述。     

火炬松成熟合子胚体胚发生与成熟试验

以火炬松成熟合子胚为外植体,开展成熟合子胚体胚诱导、胚性胚柄细胞团(ESM)维持与增殖培养及体胚成熟试验。结果表明:火炬松成熟合子胚体胚诱导率为3%,远远低于未成熟合子胚;诱导获得的多数ESM在连续的增殖培养中难以生存;不同胚性细胞系体胚发育和成熟能力不同,胚性细胞系AL-225经无植物生长调节剂培养基预培养后转入AL+ABA 50 mg·L-1+15%PEG8000培养基,体胚进一步发育,但未能获得完全成熟体胚。     

Megasporogenesis,megagametogenesis, and induction of 2n eggs with colchicine in poplar section Aigeiros

Triploid breeding through 2n eggs induction has become one of the most powerful approaches for the improvement of the genus Populus. To enhance the efficiency of triploid production in section Aigeiros, female buds (catkins) of P. × euramericana (Dode) Guinier at different developmental stages were exposed to colchicine for 2n eggs induction. The relationship between flower bud morphological characteristics (or time after pollination) and female meiotic stage (or embryo sac development) was established to guide colchicine treatments. In the resulting progeny, three triploid seedlings were obtained via embryo rescue and identified by flow cytometric analyses. Cytological observation revealed that the leptotene to pachytene stage of meiosis and 12 h after pollination might be the suitable period for 2n eggs induction with colchicine in P. × euramericana. Results and protocols related to 2n eggs induction, effective methodologies for embryo rescue, and ploidy evaluation in this study might be applicable in polyploidy breeding in section Aigeiros.     

植物三倍体培养与胚胎败育机理的研究进展

概述了人工诱导培养植物三倍体杂交的方法以及生物学方法(利用异倍体间人工杂交等)过程中胚胎败育的研究现状。分析和总结了在人工培养三倍体过程中胚胎败育的相关机理及其研究进展,并介绍了降低胚胎败育的胚抢救技术。同时简单评述了解决植物三倍体育种过程中胚胎败育研究中存在的问题,并展望了其应用前景。     

黑松合子胚和体细胞胚发育阶段的形态特征比较

为掌握黑松合子胚和体细胞胚发育进程,以未成熟合子胚为外植体,建立了黑松体细胞胚胎发生体系。对合子胚和体细胞胚发育阶段的形态特征进行观察和比较,结果表明,合子胚和体细胞胚的发育进程均可划分为8个阶段,两者同一阶段的形态特征大致相同,只有第2阶段和第4阶段有细微差别。     

异叶苦竹胚和胚乳的发育及淀粉粒动态

采用常规石蜡切片法,观察异叶苦竹胚和胚乳的发育过程,部分切片用PAS染色观察胚乳发育过程中的淀粉积累分布动态。结果表明:异叶苦竹合子的休眠期为5天,胚的发育经过二细胞原胚、多细胞原胚、梨形原胚、分化胚和成熟胚阶段,发育类型为禾本型,成熟胚具有胚根、胚芽、盾片、胚芽鞘、胚根鞘、外胚叶等典型禾本科植物成熟胚的结构。胚乳发育包括游离核、细胞化和生长成熟3个阶段,发育类型为核型。授粉后16天左右,淀粉粒开始在细胞核周围积累,以后逐渐增多,充满整个胚乳。     

LO型百合杂种后代的RAPD分析

以麝香百合系的"新铁炮"品种为母本、东方百合系的"帝伯"品种为父本,通过离体受精和胚挽救技术培育出LO型百合杂种植株LO-1,LO-2。对其进行了RAPD鉴定。结果表明:从25个随机引物中筛选出3个扩增效果较好的引物,3个引物扩增的结果表明,2个杂种后代都具有父本和母本的特征条带,并且分别具有各自的特征条带,说明杂种后代LO-1和LO-2均来自2个不同株系,是其母本与父本的杂交种。     

Somatic embryogenesis inCephalotaxus harringtonia embryo-megagametophyte co-culture

Somatic embryogenesis was initiated fromCephalotaxus harringtonia (Forbes) K. Koch embryo culture. Explants consisted of embryo and megagametophyte halves both cut longitudinally. They were removed aseptically from mature seeds and grown together on a solid Murashige and Skoog modified medium supplemented with 5 mg·l ⁻¹ 2,4-dichlorophenoxyacetic acid. Embryogenic cultures started from callus after three or more months on the primary medium. The embryogenic callus originated from the suspensor region of the embryo. All chromosome counts made in the cells of the embryonic structures demonstrated a diploid stage, which suggest that they originated from zygotic embryo tissue. The early stages of somatic embryogenic development were achieved,i.e., formation of small clusters consisting of an embryonal region made up of isodiametric meristematic cells. A more advanced stage was reached in some cultures in which the distal embryonal end of the embryo appeared smooth and opaque. The ultrastructural characteristics of the embryos, the two types of embryo cells, embryonal and suspensor cells, as well as their contents were similar to those already reported in the case of somatic embryogenesis of other conifers.     

Metabolite profiling reveals clear metabolic changes during somatic embryo development of Norway spruce (Picea abies)

Progress on industrial-scale propagation of conifers by somatic embryogenesis has been hampered by the differences in developmental capabilities between cell lines, which are limiting the capture of genetic gains from breeding programs. In this study, we investigated the metabolic events occurring during somatic embryo development in Norway spruce to establish a better understanding of the fundamental metabolic events required for somatic embryo development. Three embryogenic cell lines of Norway spruce (Picea abies (L.) Karst) with different developmental capabilities were studied during somatic embryo development from proliferation of proembryogenic masses to mature somatic embryos. The three different cell lines displayed normal, aberrant and blocked somatic embryo development. Metabolite profiles from four development stages in each of the cell lines were obtained using combined gas chromatography-mass spectrometry. Multivariate discriminant analyses of the metabolic data revealed significant metabolites (P ≤ 0.05) for each development stage and transition. The results suggest that endogenous auxin and sugar signaling affects initial stages of somatic embryo development. Furthermore, the results highlight the importance of a timed stress response and the presence of stimulatory metabolites during late stages of embryo development.     

Embryo culture is an efficient way to conserve a medicinally important endangered forest tree species Strychnos potatorum

The present study reports a protocol for germination of Strychnos potatorum (ver. Tel. Chilla) using zygotic embryo culture as an embryo rescue method. A 100% germination rate was obtained by culturing the embryos on full-strength Murashige and Skoog’s medium (MS) containing 20 g/L sucrose in comparison to McCown and Lloyd’s Woody Plant Medium (WPM). Germination rates decreased when the sucrose concentration was lower or higher than 20 g·L-1 . WPM/MS medium containing glucose at levels 30, 20, 15 g·L-1 showed a smaller percentage of germination and at quarter strength, WPM/MS medium with glucose did not respond. Multiple shoot formation was found at 1.0 2.0 mg/L BAP; 3.0 mg/L Kn; 2.0 mg/L TDZ on MS medium with 20 g·L-1 sucrose. Germination rates improved when the embryos were placed upright (vertically) in the medium. The in vitro germinated seedlings were acclimatized in a walk-in-chamber and maintained in the green house with the survival rate of 65% 75%. These plants were transferred to the field and were found to be phenotypically normal, healthy and similar to donor plants. This protocol will be useful to overcome seed dormancy and for rapid multiplication and conservation of S. potatorum using zygotic embryo culture.     

银杏种子后熟生理与内源激素变化的研究

研究银杏种子后熟期间的生理变化以及内源激素对种子休眠与萌发过程的调控作用.结果表明:银杏种胚分化不完全、发育不充分是种子休眠的主要原因,银杏种子的后熟过程实质上是种胚不断分化生长的过程.在此过程中,物质代谢、酶活性的变化均有利于种胚发育和种子发芽,种子内部经历了一系列生理生化变化去调整内源激素平衡、完成种胚发育.研究发现,内源激素在银杏种子休眠与萌发过程中起着重要的作用:GA1 3是银杏种胚后熟的关键物质;ZRs对银杏种子生长起重要的促进作用;GA1 3/ABA及ZRs/ABA相对比例的变化对种子发育、胚的后熟及种子萌发均有重要的影响.