水稻原生质体产生细胞团的冰冻保存和冻后再生植株形成

水稻(Oryza sativa L.)原生质体产生的细胞团加上10-20％的二甲亚枫(DMSO)和10-20％的蔗糖,置于液氮中保存。冻后细胞生存率达到对照的40-50％。存活的细胞在附加2×10~(-5)mol/l 2,4-D 的Linsmier-Skoog(Ls)固体培养基上再生长,然后将形成的愈伤组织块转到附加10~(-6)mol/l NAA,4×10~(-6)mol/l 激动素和10~(-6)mol/l 2 IP 及8％的蔗糖的 LS培养基上分化出芽并形成植株。     

水稻原生质体再生成熟植株

水稻原生质体培养一直是人们注意的问题,近年来这方面工作开始有了可喜的进展。本文简要报道用水稻种子成熟胚诱发的愈伤组织所制备的原生质体,经离体培养得到再生成熟植株的结果。     

水稻单倍体不定芽超低温保存和植株再生及其遗传稳定性研究

通过水稻单倍体幼穗离体培养,诱导形成了大量的不定芽。将３ｍｍ左右的不定芽转入半固体继代培养基继代培养７ｄ,而后转入１．８ｍＬ的安瓿瓶中,冰浴条件下加入预冻了的冰冻保护剂淹没材料,在冰上平衡４５￣６０ｍｉｎ后,转入程序降温仪,以１．０℃／ｍｉｎ的速率从４℃降至－４０℃,在－４０℃停留１ｈ后,投入液氮保存。将液氮保存３０ｄ左右的不定芽于３８￣４０℃的水浴中快速解冻,随即转入半固体的再生培养基,以待恢复     

高频水稻原生质体植株再生

在过去的几年中水稻(Oryza sativa L.)的原生质体培养取得了较大进展。我们在此基础上对国内大面积推广的优良粳稻品种的原生质体进行了高频植株再生的实验,并对两种不同的培养方法——琼脂糖包埋法和看护培养法进行了初步的比较研究。     

水稻悬浮细胞的超低温保存研究

本研究分析了水稻悬浮细胞的生理状态和各种冷冻前处理等因素对超低温保存后细胞存活率的影响。结果表明,继代后培养3—5天,处于对数生长期的细胞,采用二步冷冻法,超低温保存后存活率最高。电镜超微结构观察显示,0.5mol/L山梨醇预培养,10%DMSO 0.5mol/L山梨醇复合保护剂处理,液泡显著变小,数目明显减少,从而降低了细胞内自由水含量,增强细胞的抗冷冻能力。在上述合适的前处理和冷冻-化冻条件下,超低温保存水稻悬浮细胞的恢复生长率为58%,恢复生长的细胞转移到分化培养基上,可再生健壮绿苗, 移植到盆钵,在温室中长成正常结实的植株。     

Polyoxin N,a New Member of the Polyoxin Family

Eclosion hormone was purified 5,000-fold from the extracts of male adult heads of the silkworm, Bombyx mori. The fourteen-step purification procedure consisting of solvent extractions, fractional precipitations and chromatographies afforded a partially purified preparation of eclosion hormone, 1.8 μg of which showed activity in a Bombyx pharate adult. The hormone was inactivated by proteolytic enzymes. The molecular weight of eclosion hormone was estimated to be 8,400 ± 1,000 daltons by gel-filtration on Sephadex G-50.     

玉米原生质体超低温保存后芽和根的分化

植物细胞的超低温保存,已逐步发展成为一个有效的种质保存方法。近年来,随着原生质体再生植株种类的不断增多,禾谷类的主要农作物比如水稻、玉米、小麦等     

Phenotypic characterization of the progenies of rice plants derived from cryopreserved calli

The progenies of rice plants (Oryza sativa L.) differentiated from calli that had been cryopreserved and from control (non-cryopreserved) calli were used to study the influence of selection pressure during cryopreservation. The phenotypic evaluation of these progenies was based mainly on the response of seedlings and calli to freezing stress and on the characterization of protoplast and cell populations by flow cytometric analyses. The patterns of response to freezing stress, as well as the variations in some morphological and physiological cell parameters, were unrelated to the origin (cryopreserved or control calli) of the parental plants. Received: 6 August 1997 / Revised received: 28 November 1997 / Accepted: 20 January 1998     

Effects of temperature on the cell wall and osmotic properties in dark-grown rice and azuki bean seedlings

The mechanism inducing the difference in growth rate under various temperature (10–50 °C) conditions was analyzed using rice and azuki bean seedlings. The growth rate of rice coleoptiles and azuki bean epicotyls increased as temperature increased up to 40 and 30 °C, respectively, and the elongation was retarded at a higher temperature. The cell wall extensibility of rice coleoptiles and azuki bean epicotyls also showed the highest value at 40 and 30 °C, respectively, and became smaller as the temperature rose or dropped from the optimum. The opposite tendency was observed in the minimum stress-relaxation time of the cell wall. On the other hand, the cellular osmotic concentration of rice coleoptiles and azuki bean epicotyls was lower at the temperature optimum for growth at 40 and 30 °C, respectively. When rice and azuki bean seedlings grown at 10, 20, 40, or 50 °C were transferred to the initial temperature (30 °C), the growth rate of coleoptiles and epicotyls was mostly elevated, concomitant with an increase in the cell wall extensibility. The growth rate was correlated with the cell wall mechanical parameters in both materials. These results suggest that the environmental temperature modulates the growth rate of plant shoots by affecting mainly the mechanical properties of the cell wall. Electronic Publication     

植酸在水稻(Oryza sativa L.)细胞培养中的促进作用

植酸(C_6H_(18)O_(24)P_6)添加到固体和液体培养基中,能显著降低多酚氧化酶活性,稳定培养基中pH值和mV值,并能促进水稻细胞生长,增强细胞抗褐化和水渍化能力,从而改善细胞状态。在培养基中植酸的最佳添加量为0.1%,配合使用MES可以增强其稳定pH值的效果。     

从水稻花粉管分离精细胞(简报)

精细胞的分离是植物生殖工程的一个重要组成部分,是目前被子植物有性生殖研究的一个活跃领域[1,2]。随着精细胞分离技术的完善和分离出精细胞的植物类型的增加,目前对精细胞的分子生物学研究已有一些进展,主要是精细胞特异蛋白的分离[3,4]和cDNA文库的构建以及一些精细胞特异基因的分离[5,6]。     

水稻几丁质酶基因克隆RCH8的DNA结构分析

用DNA外切核酸酶Ⅲ和S1核酸酶生成连续缺失突变体,用Sanger双脱氧链终止法对该克隆进行双向DNA顺序测定,测序全长2049个碱基,初步确定了1057bp的5'端上游顺序,966bp不含内含子的完整编码区和可能的TATAbox等。所编码的322个氨基酸包括N-端20个氨基酸的信号肽,其后40个氨基酸长度的含8个半胱氨酸的hevein结构域和一个催化结构域。     

水稻马协不育与可育花药ATPase定位(简报)

马协不育系系朱英国等用农家品种“马尾粘”的不育株与“协青早选”测交和回交选育成功的。超微结构研究表明马协不育花药单核边位期绒毡层提前解体,液泡膜破裂;不育花粉外壁基粒棒少,且分布不均匀;线粒体有大的电子透明区,二核期不育花粉中央大液泡崩溃。不育花药绒毡层提前解体可能影响花粉外壁合成及花粉营养供应,大量研究表明绒毡层发育异常与花粉败育密切相关,而液泡膜的破坏会导致水解酶类的泄     

根癌农杆菌介导的水稻转基因技术体系的优化

对根癌农杆菌介导水稻遗传转化的各重要因素逐一分析,比较不同基因型、培养基成份、光照、继代次数、菌液浓度、侵染时间、干燥情况等因素对遗传转化的影响。通过培养条件的优化,建立了一个高效的水稻转化技术体系。研究结果表明,相比较于暗培养,光照培养进行愈伤诱导可以使诱导天数缩短4~5d;诱导培养基中加入适量的激素可以使籼稻愈伤诱导提高25%~30%;分化培养基中加入适量的氨基酸和甘露醇可以使植株再生频率提高15%~20%。