不同基因型小麦花药培养过程中药壁变化的研究

在相同的培养条件下,比较了不同基因型小麦花药培养过程中药壁变化的差异。组织切片观察表明：易诱导材料花药培养过程中,药壁细胞发育充实,活力强,降解衰退较难诱导材料有所延缓;两种材料的花药刚离体时绒毡层就存在着明显的差异。     

花药培养中花粉去分化启动条件和药壁变化的初步研究

研究了对花粉去分化启动的必需外源因素和花药培养过程中的药壁变化。结果如下:1.对于花药培养中的花粉去分化启动,至少要有糖的外源供应。2.花粉对等分裂的高峰出现在颤绒层消失之后,这暗示颤绒层的适时衰退可能有助于花粉的去分化启动。3.花药培养过程中,在药壁会出现糖和淀粉的积累,表明药壁还有若干合成活动。4.材料不同、培养基不同,药壁颤绒层衰退的迟早和药壁可溶性蛋白带谱的变化不同,这可能暗示材料差异和培养基对花药培养效率的影响,部分是通过影响药壁状况而实现的。     

低温预处理对水稻花药培养中花药壁褐变的结构影响

药壁褐变是影响花药离体培养效率的因素。研究了粳稻品种台中6(5OryzasatvaL.)花药培养前低温预处理对药壁褐变的影响,观察了花药褐变前后药壁各层的变化及褐变对小孢子发育的作用。结果表明:药壁褐变主要发生在表皮与药室内壁。10℃低温预处理在花药培养前期可有效延缓表皮与药室内壁膜结构的降解速度,减缓褐变发生;花药经低温处理后,药壁中层细胞膨大,绒毡层降解速度减缓,有利于花粉脱分化。药壁褐变会影响药腔内小孢子的活力和继续发育,但不是制约花粉愈伤组织形成的关键因素。     

油菜花药培养研究(初报)

试验了不同培养基配方和不同的激素及附加成分对油菜花药培养的影响。结果表明,对于诱导油菜花药形成愈伤组织,Miller和B_5,配方较好。但在大多数情况下,愈伤组织易从花丝及药隔组织形成。在MS培养基中加入适量的BA和NAA以诱导芽的分化,明显优于用激动素及IAA的组合。诱导培养基的组或对形成的花药愈伤组织随后的分化有明显的影响。温度是提高小苗移栽成活率的关键因素。由甘蓝型油菜×白菜型油菜的F_1植株的花药培养诱导形成的部分植株中出现了明显的分离现象。对花药培养诱导形成的植株的第二代群体的观察表明少数植株的后代群体表现整齐一致。这些结果说明在油菜的花药培养中可能有少数花粉植株的形成。讨论了油菜花药培养中存在的问题。     

甘蓝花药培养胚状体诱导形成影响因子研究

用甘蓝F1、F2和自交系S33个世代6种基因型材料进行甘蓝花药培养诱导胚状体形成影响因子研究。结果显示:(1)高浓度蔗糖对甘蓝胚状体形成具有显著的诱导作用,6%蔗糖浓度是甘蓝花药培养的最适浓度,其胚状体的诱导率最高达12.2%;(2)材料基因型是影响花药培养的主要因素,F2和F1代材料胚状体诱导效果好,且胚状体诱导率F2代(F2P192和F2P194)18.9%比F1代(F1S17和F1S13)17.1%较高,但差异不显著,自交系S3代材料很难诱导出胚状体;(3)B5培养基比MS培养基更适合甘蓝花药胚状体的诱导培养。结果表明,甘蓝F2代是其花药诱导培养胚状体的最佳基因型材料,B5 2.0 mg/L 2,4-D 2.0 mg/L KT 6%Suc是甘蓝花药诱导培养胚状体的最适培养基。     

高温预处理对大白菜游离小孢子培养的效果(简报)

自从Keller在甘蓝型油菜花药培养中首次用高温处理提高了花粉胚的诱导频率以来,该方法被广泛用于芸苔属不同作物的花药培养     

在低温预处理及花药培养期间高氯酸汞处理对大麦花药生产力的影响

在春大麦(sabarlis品种)幼穗低温预处理过程中,使用乙烯的吸收剂高氯酸汞溶液处理,明显提高花药培养中花药的诱导率,以及每个花药形成的愈伤组织数目。在花药培养中,此种处理仅增加花粉愈伤组织的形成,而对花药的诱导率影响不明显。     

小麦花粉植株的遗传学研究

自1971年4月我们用花药培养方法诱导出小麦单倍体花粉植株以来,国内外学者在小麦花药培养研究中作了大量工作,诸如:离体条件下的雄核发育的研究,提高诱导花粉植株频率的各种因素的研究,其中包括培养基的改进、材料基因型差异在诱导中的作用和培养条件,以及供取花药植株的环境条件的研究等。本文拟在多年研究的基础上,对小麦花粉植株的遗传学表现,包括目前大家所关心的花粉植株的生活力,以及花粉植株的变异等问题做进一步的探讨。     

低温预处理影响籼稻花药培养效率的研究

低温预处理使籼稻花药在培养初期退化花粉百分率低于对照;多核花粉百分率明显高于对照。经过低温预处理的花药来见新的可溶性蛋白谱带出现;淀粉酶同工酶谱带少和弱于对照,过氧化物酶同工酶中碱性区活性初期弱而后期强于对照。低温处理使药壁组织衰退进程有所延缓,但未能制止衰退,也未出现如粳稻中所见的药壁细胞充实发育的现象。低温预处理只能稍微延缓籼稻药壁组织的衰退进程,有限度地促进籼稻花药培养效率。     

小麦花粉植株诱导条件的研究

根据近几年来国内外有关植物花药培养方面的主要资料,我们进行了小麦花粉植株诱导条件的实验研究。除培育出一批花粉植株外,还观察了小麦花粉植株诱导过程中各种培养基成分对于花粉愈伤组织形成和分化过程的影响程度,比较了各种实验材料之间的差异,讨论了与培养过程有关的光照、温度等问题。其中除部分1973年材料已在《遗传学通讯》发表外,再根据1973、1974年两次实验结果作简要总结,以便取得经验,不断改进培养技术和方法,提高培养效率,为小麦花粉植株的培育,为小麦单倍体育种工作打下良好的基础。     

Anther wall layers control pollen sugar nutrition inLilium

Summary Anthers ofLilium were for the first time investigated at the ultrastructural level in order to appreciate the possible ways of sugar transport in the microsporangium. Our results have shown that the cells of the outer anther wall layers and the cell of the connective were interconnected by plasmodesmata, thus allowing assimilates to travel through the symplasmic pathway from the vascular bundle to the most internal middle layer (ML 1). ML 1 was devoid of cell communication throughout pollen development. Tapetal cells were also lacking plasmodesmata on their external face towards ML 1, but adjacent tapetal cells developed lateral junctions: the tapetum could represent a syncytium. Sugars destinated to pollen in the loculus have then to cross the ML 1 and the tapetal layers by the apoplasmic pathway; it is suggested that these two envelopes could be involved in the control of sugar transport from the outer anther wall layers to the locular fluid. Before microspore mitosis, the tapetum degenerated but ML 1 remained structurally unchanged. During pollen development, the guard cells of stomata were lacking cell communication, and preserved their starch content, which could be the sign of photosynthesis within the anther wall. In order to check whether these structural disconnections in anther tissues corresponded to physiological barriers, isolated pollen and stamens were cultivated during the anther maturation phase, on a medium containing increasing concentrations of sucrose (0 M, 1/6 M, 1/2 M, 1 M). After 7 days of culture, isolated pollen was engorged with starch grains and was unable to germinate, whereas in cultivated stamens, pollen did not contain any starch grain: sporophytic tissues, however, accumulated abnormal amylaceous reserves. These results strongly suggest that the anther wall layers, in particular ML 1, starve pollen with sugars during its maturation. They are acting as a physiological buffer storing nutriment surplus in starch grains.Abbreviations ML 1 middle layer 1 - ML 2 middle layer 2 - PAS periodic acid Schiff - PATAg periodic acid thiosemicarbazide silver nitrate     

Anther Factor(s) in Barley Anther Culture

The effects of anther tissues were studied systematically on microspores forming multicellular units and furthermore on pollen callus formation in the anther culture of Hordeum vulgare (cv. Sabarlis). Anther productivity was found to be greatly enhanced by use of medium previously conditioned by anthers. In 15 experiments observed, anthers produced 26 times on average more calli in the conditioned medium than in control, in a few cases, even more than 80 times more calli formed. According to this, the authors supposed that cultured anthers released some components, anther factor (s) (AF), which is important to androgenesis in the culture. To achieve high yields of callus, culture was restricted to anthers which had been subpected to cold pretreatment. The temperature stress could not be replaced by the AF. However, for conditioning medium, anthers at binuclear stage were found to be more effective than the test anthers either with or without the pretreatment. Anthers from other 8 barley varieties were also effective for conditioning, as the difference of anther productivity still existed in the culture with conditioned medium between various genotypes tested. Anther response and callus yield were increased in both the culture of anthers at mid and late-uninuclear stage by use of conditioned medium. AF interacted synergistically with m-inositol. Cytological observation showed that AF increased apparently the formation of MPGs, while m-inositol mainly stimulated callus formation from MPGs. To some extent, the effect of exogenous hormone(s) could be replaced by AF. The anther response and pollen callus yield could be much enhanced by increasing anther inoculation density, which also raised the AF level in the culture. Thus, by use of the temperature stress prior to anther culture and culture of test anthers in conditioned medium with m-inositol, or at higher inoculation density, a very high production of pollen callus could be obtained in barley anther culture. For meeting the more specialized requirements of less responsive species or genopypes, the principles given here may be provide some basic information.     

花药培养过程栽培稻花粉不育杂种F1与亲本台中65的细胞学研究

利用活体压片、半薄及超薄切片技术,对栽培稻(Oryza sativa L.)花粉不育杂种F1及育性正常的亲本台中65离体培养前后的小孢子及花药壁进行细胞学研究.结果表明:与台中65相比,杂种F1的花粉小孢子在发育至单核中-晚期,出现比例较高的胞质凝聚小孢子和少量星型小孢子,正常小孢子和淀粉化小孢子比例降低.在离体条件下,胞质凝聚小孢子、星型小孢子、正常小孢子、液泡化小孢子、淀粉化小孢子的发育主要沿着胞质凝聚败育过程、孢子体发育过程、配子体发育过程、液泡化过程和淀粉化过程进行;药壁组织在离体条件下,杂种F1比台中65的绒毡层降解速度快,中层膨大程度高.杂种F1与台中65在离体培养下小孢子发育及药壁细胞学的差异主要是受到S-a座位内等位基因互作及离体培养环境的影响.     

Uitrastructure of Anther Tissue and Microsporogenesis in Tai Gu Nuclear Male-Sterile Wheat

This paper deals with the electron microscopic observations of anther tissues and microsporogenesis of Tai Gu male sterile wheat plant, which is a new species initially discovered in Tai Gu nE Shansi province, China. The results are summarized as follows: 1. There was a tendency of early development of tapetum cells in Tai Gu sterile wheat plant, i.e. the tapetum cells seemed to have developed rapidly toward maturation between the period of pollen mother cell formation and prophase I of reduction division, which is followed by degeneration. Its ultrustructure is somewhat similar to the highly developed tapetum cells of fertile plant at the time of development from tetrad to early microspore stage. A significant character of the nltrastructure of the highly developed mature tapetum cells was the appearance of many sharply-defined endoplasmic reticulum, which demonstrated in a parallelly arranged structure in section. Phagocytic vacuoles were often surrounded by the endoplasmic reticulum. 2. At the prophase I of reduction division, the ultrastructure of pollen mother cells in sterile plant began to show some disparities from that of fertile pIant. Among which there were some concentric circle-shaped or bundle-shaped endoplasmic reticula and many ribosome aggregation in the pollen mother ceils of sterile plant. Perhaps, these features might be relevant to the break down of microtrabecular system. In addition, some large or small dark osmiophilic particles were visible in the cells. From all the feature observed, the fact that activities of endoplasmic reticulum in the anther of Tai Gu wheat is relation to protein synthesis warrants our attention.     

林木花药培养研究进展及展望

多数林木在遗传上高度杂合,给育种和遗传研究带来很多不便。与农作物相比,林木花药培养的意义更为重大。对国内外林木花药培养的现状及影响花药培养的主要因素进行了概述,讨论了其在21世纪分子生物技术时代的应用前景,旨在促进林木花药培养技术的完善并开拓其应用领域。     

红菜薹雄性不育系花药败育的细胞形态学观察

采用石蜡切片技术,在光学显微镜下系统研究了红菜薹(Brassica campestris L.ssp．chinensis L.var．utilis TsenetLee．)波里马胞质雄性不育系(Polima CMS)、红菜薹萝卜胞质雄性不育系(Ogura CMS)及相应保持系花药发育过程的细胞形态学特征。观察结果表明：红菜薹Polima CMS花药发育受阻于孢原细胞阶段,不形成花粉,属无花粉型,此不育系花药不形成绒毡层和中层;而红菜薹Ogura CMS花药败育发生于小孢子母细胞期或四分体时期,表现为绒毡层细胞异常,挤压四分体,导致四分体和绒毡层同时解体而败育。     

玉米花药发育的细胞生物学与分子遗传学的研究方法

雄性不育技术在玉米杂种优势利用和杂交种生产中发挥着重要作用,玉米花药发育和雄性不育的细胞生物学与分子遗传学研究是雄性不育技术利用的前提和基础。玉米花药发育是一个复杂的生物学过程,需要孢子体基因与配子体基因的协同表达调控。从玉米花药的形态结构、花药发育时期的划分、花药败育类型、花药发育的细胞学研究方法、花药发育的组学研究方法、花药发育分子遗传学研究方法等方面进行综述,以期为玉米核不育机制研究与雄性不育技术产业化应用提供方法学指导。     

盾叶薯蓣花药培养及单倍体植株的获得

以处于单核期的盾叶薯蓣花药为材料,经愈伤组织途径成功获得单倍体盾叶薯蓣新材料。对盾叶薯蓣花药培养、植株再生过程的研究表明:来自不同居群的外植体对花药愈伤组织诱导有显著影响;W14是适合花药愈伤组织诱导的基本培养基;花药愈伤组织增殖和分化的适宜培养基为:MS基本培养基 BA2.0mg/L IAA0.2mg/L;生根培养基为添加了IBA2.0mg/L、NAA0.4mg/L和0.5g/L活性碳的MS培养基。以流式细胞仪和根尖染色体压片法检查花培植株的结果表明,有8%～12%的个体为单倍体。实验结果为进一步开展单倍体育种及相关理论研究提供了材料和技术基础。     

Anther structure and pollen development in Melicoccus lepidopetalus (Sapindaceae): An evolutionary approach to dioecy in the family

Anther and pollen development in staminate and pistillate flowers of dioecious Melicoccus lepidopetalus (Sapindaceae) were examined by light and electron microscopy. Young anthers are similar in both types of flowers; they consist of epidermis, endothecium, two to four middle layers and a secretory tapetum. The microspore tetrads are tetrahedral. The mature anther in staminate flowers presents compressed epidermal cells and endothecium cells with fibrillar thickenings. A single locule is formed in the theca by dissolution of the septum and pollen grains are shed at two-celled stage. The mature anthers of pistillate flowers differ anatomically from those of staminate flowers. The epidermis is not compressed, the endothecium does not develop fibrillar thickenings, middle layers and tapetum are generally persisting, and the stomium is nonfunctional. Microspore degeneration begins after meiosis of microspore mother cells. At anthesis, uninucleate microspores and pollen grains with vegetative and generative nuclei with no cytokinesis are observed. Some pollen walls display an abnormal exine deposition, whereas others show a well formed exine, although both are devoid of intine. These results suggest that in the evolution towards unisexuality, the developmental differences of anther wall tissues and pollen grains between pistillate and staminate flowers might become more pronounced in a derived condition, such as dioecy.