感染病毒的番茄花药发育过程中游离脯氨酸含量与花粉萌发率的变化(简报)

植株染病毒的番茄花药发育过程中游离脯氨酸积累开始较晚，任一花药发育时期花药中的游离脯氨酸含量均低于同一发育时期的健康植株，花粉萌发率也低。     

植物花粉培养研究进展

花粉培养又称为游离小孢子培养,指将发育到一定阶段的花粉从花药中游离出来成为分散或游离状态,通过培养使花粉粒脱分化,进而发育成完整植株的过程。花粉培养的主要目的是获得单倍体植株,进而得到双单倍体(double haploid,DH)植株,最终获得纯合系物种。本文对花粉培养形成植株的物种信息进行了收集整理,概述了国内外花粉培养的一些最新研究进展,包括影响花粉培养形成胚的因素以及提高花粉胚产量的措施,并对花粉培养的前景进行了展望。     

植物花粉培养研究进展

花粉培养又称为游离小孢子培养,指将发育到一定阶段的花粉从花药中游离出来成为分散或游离状态,通过培养使花粉粒脱分化,进而发育成完整植株的过程。花粉培养的主要目的是获得单倍体植株,进而得到双单倍体(double haploid,DH)植株,最终获得纯合系物种。本文对花粉培养形成植株的物种信息进行了收集整理,概述了国内外花粉培养的一些最新研究进展,包括影响花粉培养形成胚的因素以及提高花粉胚产量的措施,并对花粉培养的前景进行了展望。     

超低温保存的梅花花药萌发率和授粉后的结实能力

试验比较梅花花粉和花药超低温保存后的花粉萌发率、授粉结实率和座果率的结果表明:(1)超低温保存花药用室温、自来水冲洗和温水浴3种方法化冻后的花粉萌发率差异不显著。(2)花药超低温保存1h后的花粉萌发率与新鲜花粉差异不显著。(3)超低温保存1个月和1年的花粉和花药都有授粉结实能力。     

水稻游离花粉培养诱导形成绿色植株及有关作用因素的研究

游离花粉粒培养诱导产生植株的研究,近几年来在几种茄科植物上获得成功。但在禾本科植物上迄今未见有成功的报道。1978年我们从水稻花药预培养4—7天的花粉培养诱导出白化植株。1979年这一研究又获得进展,从预培养2天和2天以上的花药分离出的花粉都成功地诱导出绿色植株。在此同时,我们还对一些影响花粉启动、发育和愈伤组织分化的因素进行了研究,结果报道如下。     

水稻花药和花粉液体培养的研究

对四个粳稻品种、两个粳籼杂种F_1的花药进行漂浮培养,几天后,大量花粉由花药自然散落到培养液中。以后无论是这些悬浮在培养液中的花粉或仍包含在花药中的花粉都能形成大量愈伤组织,并进一步分化为小植株。这一培养方法不仅可以大幅度提高水稻花粉植株的诱导率,并为在细胞水平上进行遗传饰变的研究提供良好的实验体系。     

TA29—Barnase嵌合基因导入对烟草花药绒毡层及花粉发育的影响

比较研究了烟草(Nicotiana tabacum L.)TA29-Barnase转基因不育植株和正常植株的花药绒毡层及花粉发育的全过程。研究表明,外源基因在花药中特异表达导致绒毡层细胞的提前降解,这种降解一般在减数分裂早期开始,至四分体时期完成,而正常花药绒毡层的降解发生在二细胞雄配子体初期,至花粉发育的后期方才完成。转基因植株花药绒毡层的降解在细胞结构上表现为:最初发生细胞的液泡化,然后细胞核凝聚,最后整个细胞溃解。转基因植株的花粉母细胞则在减数分裂过程中逐渐降解、退化,只有少数花粉母细胞能够顺利完成减数分裂发育成小孢子。观察结果还表明外源基因在花药中的表达是不均一的。对转基因不育和自然败育在细胞结构上的不同表现进行了讨论。     

菠萝蜜花药发育及花粉萌发研究

研究花药发育过程和花粉萌发条件是菠萝蜜稳产优质的基础。采用石蜡切片和离体培养的方法,对菠萝蜜花药的发育和花粉萌发进行了研究,结果表明：菠萝蜜的花药有4个花粉囊;减数分裂的胞质分裂有连续型和同时型两种形式,形成了等双面体排列和四面体排列的四分体;成熟花药的表皮细胞积累有大量的单宁。160 g·L^-1的蔗糖和0.25 g·L^-1的硼酸混合溶液对菠萝蜜花粉萌发具有明显的促进作用,CaCl₂对菠萝蜜花粉萌发作用不明显。     

温度对桃离体花药散粉及花粉萌发的影响

以目前生产栽培较多的‘湖景蜜露’、‘霞晖6号’和‘白凤’3个桃品种为试材,连续2年调查了不同温度处理下花药失水率、花药散粉时间以及花粉离体萌发特性等变化。结果表明：桃花药于相对低温条件下散粉失水率较低,随散粉温度升高失水率相应上升;花药开裂所需时间与处理温度呈相反趋势;3个品种花粉离体萌发率随散粉温度的升高而下降。离体花药在超过30℃的温度条件下散出的花粉在萌发过程中出现花粉管变短、花粉瘪小的概率增多的现象,表明高温促使花药脱水和散粉加快,但降低了花粉活力。在桃树花期以及制备花粉时外界环境温度应控制在30℃以下。     

鸡蛋花不育机理初步研究

本文探索鸡蛋花开花不结实的原因。通过对鸡蛋花雌、雄性器官的切片观察和离体花粉萌发,发现其花粉和胚囊均正常,离体花粉可萌发花粉管;但鸡蛋花的花筒长约1 cm,花筒基部湿度较高;开花时,雌蕊柱头和雄蕊花药均不能伸出花筒,花药不能开裂释放花粉,产生了传粉障碍,最终导致不育。     

开花期低温胁迫对水稻花粉性状及剑叶理化特性的影响

以耐冷水稻品种996和冷敏感品种4628为材料,开花期在人工气候室进行7 d低温（06:00-8:00和19:00-23:00,19 ℃;08:00-10:00和16:00-19:00,21 ℃;10:00-16:00,23 ℃;23:00-06:00,17 ℃）和适温（06:00-8:00和19:00-23:00,24 ℃;08:00-10:00和16:00-19:00,26 ℃;10:00-16:00,30 ℃;23:00-06:00,22 ℃)处理,研究了低温胁迫对水稻花药开裂、花粉性状及剑叶叶绿素、可溶性糖和可溶性蛋白质含量、膜透性等理化特性的影响.结果表明：开花期低温胁迫导致水稻花药开裂系数、花粉萌发率显著下降,中部和下部颖花的不育花粉率增加.996的花药开裂系数和花粉萌发率显著高于4628,表明耐冷品种996在开花期低温胁迫下能保持较好的花粉散落特性和花粉萌发特性.低温胁迫下,996的剑叶可溶性蛋白质和游离脯氨酸含量及其增幅显著高于4628,而丙二醛含量和相对电导率及其增幅却显著低于4628,说明耐冷品种在低温胁迫下的保护性反应更迅速和强烈,其膜结构及功能更稳定.     

Fructokinase and hexokinase from pollen grains of bell pepper (Capsicum annuum L.): possible role in pollen germination under conditions of high temperature and CO2 enrichment

The processes of pollen grain development and germination depend on the uptake and metabolism of pollen sugars. In pepper (Capsicum annuum L.), initial sugar metabolism includes sucrose hydrolysis by invertase and subsequent phosphorylation of glucose and fructose by hexose kinases. The main objective of this study was to investigate changes in fructokinase (EC 2.7.1.4) and hexokinase (EC.2.7.1.1) activities in pepper flowers during their development, and to study the possible roles of these enzymes in determining pollen germination capacity under high temperature and under CO(2) enrichment, previously shown to modify sugar concentrations in pepper pollen (Aloni et al., 2001 Physiologia Plantarum 112: 505-512). Fructokinase (FK) activity was predominant in pepper pollen, and increased during pollen maturation. Pollen hexokinase (HK) activity was low and did not change throughout pollen development. High-temperature treatment (day/night, 32/26 degrees C) of pepper plants reduced the percentage of pollen that germinated compared with that under normal temperatures (26/22 degrees C), and concomitantly reduced the activity of FK in mature pollen. High temperature also reduced FK and HK activity in the anther. Under high ambient CO(2) (800 micro l l(-1)) pollen FK activity was enhanced. The results suggest that pollen and anther FK may play a role in the regulation of pollen germination, possibly by providing fructose-6-phosphate for glycolysis, or through conversion to UDP-glucose (UDPG) to support the biosynthesis of cell wall material for pollen tube growth. High temperature stress and CO(2) enrichment may influence pollen germination capacity by affecting these pathways.     

Possible correlation between high temperature-induced floret sterility and endogenous levels of IAA,GAs and ABA in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

In order to explore the possible physiological mechanism of high temperature induced sterility in rice, we examined the floret sterility and endogenous plant growth regulator contents in pollens of two hybrid rice cultivars Shanyou63 and Teyou559 that are tolerant and susceptible to high temperature, respectively. Indexes of floret sterility, pollen activity, and variation of endogenous indole-3-acetic acid (IAA), gibberellic acids (GAs), abscisic acid (ABA), free proline and soluble proteins in anthers were measured. We found that during the course of high temperature treatment, both cultivars exhibited a marked decrease in pollen activity, pollen germination and floret fertility; however, the high temperature tolerant Shanyou63 showed a much slower rate of decrease than the high temperature susceptible Teyou559. In addition, anthers of both cultivars displayed a decrease in the contents of IAA, GAs, free proline and soluble proteins but an increase in the ABA content. Yet compared to Teyou559, Shanyou63 retained significantly higher levels of free praline and GAs and a lower level of ABA, along with higher pollen vigour and pollen germination rate even after prolonged high temperature treatment. Our study suggests a possible correlation between pollen viability/floret sterility and high temperature-caused changes in IAA, GAs, ABA, free proline and soluble protein contents. The severity in these changes may reflect the variation of rice cultivars in their heat stress sensitivities for floret development.     

辣椒雄性不育系与保持系花药POD、SOD和EST同工酶表达差异研究

利用辣椒质核互作雄性不育材料,采用聚丙酰胺凝胶垂直板电泳技术,研究了在花药发育不同时期,甜椒和辣椒核质互作雄性不育系及其保持系POD、SOD和EST同工酶的差异.结果表明:不育系与保持系,在花药发育的不同时期POD同工酶表达种类及表达活性之间存在差异,其中甜椒不育系在减数分裂期POD同工酶的活性非常低,辣椒雄性不育系在减数分裂期及花粉成熟期分别多一条POD特征带.不育系与保持系之间SOD同工酶的表达活性及其表达种类无明显差异.在减数分裂期和花粉成熟期,甜椒和辣椒不育系与保持系花药EST同工酶酶带均存在明显的差异.     

Studies on the Free Amino Acid Content in Anthers of Malesterile and Fertile Plants of Taigu Wheat,with Special Reference to Free Proline Content

Biochemical analyses were made on anthers and pistils at various developmental stages of both male-sterne and fertile plants of Taigu wheat. Analyses ineluded total free amino aeids and free proline. The following results were obtained: 1. There was no significant difference between the content of free proline in anthers of male-sterile and fertile plants at reduction division of mierospore mother cells. 2. In anthers with early uninucleate miorospores, the content of free proline of fertile plants was remarkably higher than that of male-sterile plants. It is interesting to note that at this stage the content of free proline in fertile plants rose to 1.65% of the dry weight of the anther, constituting 50% of the total free amino acids, and amounted to 7-fold of that in male-sterile plants. This result is in line with the results obtained with most cytoplasmic malesterile plants reported by other workers, although malesterility in Taigu wheat is controlled by the nueleus. 3. In pistils, at the stages eorresponding to the early uninneleate and the binueleate stages of the pollen, the free proline content of fertile plants was twice as much as that of the male-sterile plants. This differenee disappeared gradually after fertilization. 4. Tile content of total free amino aoids did not fluetuate as much as the free proline content. There was no differenee in anthers of both types of plants during reduction division of mierospore mother cells. In anthers with early uninueleate pollen grains, total free amino acid content of fertile plants exceeded that of male-sterile plant, the difference levelled off at latter stages. In pistils, before fertilization, the content of total free amino acids of the fertile phmts was slightly higher than that of the male-sterile plants. After fertilization t}fis difference was no nmre noticeable.     

缙云山四川大头茶花粉游离脯氨酸含量与生殖产量特征研究

四川大头茶（Gordonia acuminata）是缙云山亚热带常绿阔叶林的优势树种之一,在缙云山的很多群落类型中均有分布。对缙云山四川大头茶的花粉游离脯氨酸含量的影响因素以及花粉游离脯氨酸含量与四川大头茶生殖产量之间的关系进行了研究。研究表明,缙云山四川大头茶的花粉游离脯氨酸含量不受植株年龄大小的影响,但与植株所处的群落类型有很大关系。常绿阔叶中四川大头茶的花粉游离脯氨酸含量最高,四川大头茶纯林中次之,针阔混交林中四川大头茶花粉游离脯氨酸含量最低。四川大头茶花蕾产量和果实产量随植株年龄的增大而增大,但座果率与植株年龄大小无关。对花粉游离脯氨酸含量与花蕾产量、果实产量和座果率之间的相关关系分析,花粉游离脯氨酸含量与花蕾产量不具有相关关系,与果实产量具有正相关关系,与座果率具有极显著正相关关系。这证实了与受精作用无关的生殖产量特征不受花粉游离脯氨酸含量影响,而与受精作用有关的生殖产量特征将受花粉游脯氨酸含量影响的研究假设。由于与花粉游离脯氨酸含量呈极显著正相关关系,座果率具有和花粉游离脯氨酸含量一样的群落间变化模式。     

Trihalogenated methylsulfonamides as specific male gametocides

A new class of male gametocide is described: trihalomethylsulfonamides, the most active example of this class being trifluoromethylsulfonamide (TFMSA). TFMSA induces male sterility, specifically, without detectable effects on other plant functions. Male sterility induction in maize (Zea mays, gaspe flint) required minimally 200 g TFMSA per plant and this rate was used in a metabolite sampling time-course experiment to determine the earliest detectable change in metabolites of developing florets. Metabolites profiled were amino acids, callose, fatty acids, flavones, phenylpropenoids, sporopollenin and starch, all of which are related to successful pollen development. Changes in proline and starch were the earliest statistically significant differences observed between florets of control plants and TFMSA-treated plants. These metabolic differences were observed before symptoms of pollen failure were evident. In subsequent experiments, transient increases in glume proline and decreases in anther proline were linearly related to sub-effective rates of TFMSA (0, 25, 50, 75 and 100 g plant^–1). Increases in glume proline faded during development whereas decreases in anther proline linearly related to TFMSA rate became more prominent. Changes in all other metabolites profiled were not linearly related to TFMSA rate. Related experiments showed that florets from TFMSA-treated plants were not capable of converting ¹⁴C-glutamate to ¹⁴C-proline, and the anther transport capacity of ¹⁴C-proline in TFMSA-treated plants was significantly reduced. It is inferred that TFMSA induces male sterility by interfering with the transport of proline from the site of synthesis to the site of accumulation, resulting in feedback inhibition of proline biosynthesis, ultimately starving the developing anther of proline.     

棉花洞A型核雄性不育系花药败育过程中的生化变化

研究了陆地棉洞A型核雄性不育系(抗A1,MA)不育和可育花药不同发育时期可溶性碳水化合物、游离氨基酸和IAA、GA3、ABA含量的动态变化。结果表明,不育花药中可溶性糖含量偏高,缺乏淀粉积累;有4种游离氨基酸含量在可育与不育花药问存在显著差异,其中不育花药天门冬氨酸含量偏高,这可能是其败育的原因之一,而脯氨酸、精氨酸、苯丙氨酸含量异常,则是不育花药败育的结果;在花药主要败育时期之前,不育花药中ABA含量极显著偏高,IAA、GA3含量极显著偏低。不育花药中IAA、GA3、ABA含量变化与洞A型核雄性不育系花药败育密切相关。     

雄性不育无籽刺梨花药发育过程中生理特性的初步研究

为探究无籽刺梨(Rosasterilis)雄性不育原因,采用1%I2-KI染色法观察花粉活性,并对无籽刺梨和正常可育刺梨(R.roburghii)花药不同发育时期的生理生化指标进行了研究。结果表明,无籽刺梨的败育花粉占95.5%,刺梨的正常花粉占99%。刺梨花药的可溶性蛋白质、可溶性糖和脯氨酸含量在各时期的总体变化趋势相似,可溶性淀粉含量呈上升趋势,而无籽刺梨花药的可溶性蛋白质、可溶性糖、淀粉和脯氨酸含量在各时期的变化无规律可循,且花粉成熟期这4种物质的含量均明显低于刺梨,即花粉成熟期缺少各营养物质的积累。在花药发育过程中,无籽刺梨的SOD活性均低于刺梨;MDA含量呈上升趋势,且上升幅度比刺梨大;MDA含量和POD活性均高于刺梨。因此,营养物质的匮乏和酶系统的紊乱可能是导致无籽刺梨雄性不育的原因。     

太谷核不育小麦花药内游离脯氨酸和总氨基酸含量的变化及其与育性的关系

本文对显性单基因控制的太谷核不育小麦不同发育阶段的可育株和不育株的花药及雌蕊内游离肺氨酸和游离总氨基酸的含量进行了分析。结果表明:(1)在小孢子母细胞减数分裂期,不同育性花药之间游离脯氨酸的含量无明显差异,且含量较低。(2)在小孢子单核初期,可育花药内游离脯氨酸的含量显著高于不育花药,是不育花药的7倍,比减数分裂期增加20倍,高达其干重的1.65％,占其游离总氨基酸的50％。(3)在雌蕊中,游离脯氨酸的含量远远低于花药,不同育性植株之间差异不很明显。(4)关于游离总氨基酸的含量,在花药中减数分裂期,不同育性植株之间无明显差异;在小孢子单核初期,可育株高于不育株。在雌蕊中,相应于小孢子单核初期时,可育株稍高于不育株,受精后迅速趋于一致,但整个变化幅度不大。