毛钩藤的小孢子发生和雄配子体发育

在光学显微镜和透射电镜下观察了毛钩藤(Uncaria hirsuta Havil.)的小孢子发生和雄配子体发育过程.结果表明,毛钩藤花两性,具5枚雄蕊,花药4室,花药壁由表皮、药室内壁、中层和绒毡层组成,花药开裂时,药室内壁高度纤维化带状加厚.花药壁的发育方式属于双子叶型,小孢子母细胞减数分裂的胞质分裂为同时型.小孢子在四分体时期开始沉积花粉外壁,小孢子大液泡化时期开始沉积花粉内壁.成熟花粉为2-细胞型.毛钩藤的花粉发育特征和茜草科植物基本一致.毛钩藤绒毡层属于分泌型,双重起源,分别起源于次生周缘层和药隔细胞.小孢子发育早期绒毡层开始降解并分泌形成大量乌氏体,花药开裂时绒毡层完全消失,剩下少量乌氏体.小孢子早期内壁加厚突出形成,小孢子细胞核分裂以后内壁加厚开始脱落,花药开裂时,只剩下少量的内壁加厚突出.初步推测,内壁加厚突出与乌氏体共同作用为雄配子体的发育提供营养物质.     

温敏雄性不育水稻培矮64S花药发育过程中钙的变化

采用焦锑酸钾沉淀法研究了温敏雄性不育水稻(Oryza sativa L.)培矮64S在高温引起雄性不育与正常可育花药发育过种中Ca2+的分布变化.结果表明,当培矮64S生长在较高温度条件下引起雄性不育,与可育花药相比,不育花粉母细胞中有较多的液泡、较多的Ca2+沉积和较少的线粒体,并且有较多的Ca2+沉积在不育花药的中间层、表皮层和绒毡层中.到四分体与单细胞花粉时期,不育花药的木质部细胞的次生加厚壁上有较多的Ca2+沉淀,连接组织中的Ca2+沉淀也大大增加,所有不育花粉外壁较厚而发育都不正常.在单核细胞早期,不育花粉的四分体细胞中有较明显的大液泡出现.不育花药中的Ca2+在花药发育的各时期均比可育花药要多.这些结果说明在高温生长条件下,花粉母细胞发育的异常、花药中Ca2+沉积的增加、绒毡层与花粉外壁发育的异常可能与培矮64S花粉败育相关.     

水稻红莲型细胞质雄性不育系小孢子不同发育时期花药总蛋白质比较分析

采用双向凝胶电泳对水稻红莲型细胞质雄性不育的不育系小孢子发育单核期和二核期花药总蛋白进行了分离,通过银染显色,获得了分辨率和重复性较好的双向电泳图谱,且单核期和二核期花药总蛋白质在双向电泳胶上分布的图谱十分相似。PDQuest 2DE图像分析软件在等电点(pI)3.0~10.0、分子量(M.W.)9.0~98.0 kD之间可识别约1 800个蛋白质点。比较分析发现单核期和二核期花药中共有241个差异表达的蛋白质点,其中仅在单核期中表达的点数为125,仅在二核期中表达的为13点;表现为表达量差异的105点,其中在二核期表达下调的点数为70点,表达上调的为33点。还对蛋白质点集中的区域(pI 4.5~8.0,M.W.25.0~70.0 kD)中的41个差异蛋白质点进行了分子量和等电点分析。     

水稻花药总蛋白质双向电泳方法的改良与优化(简报)

双向电泳作为蛋白质组学核心技术之一,目前已广泛地应用在植物领域,并且成功应用于水稻代谢和调节等方面的研究。谢锦云等利用溶液法提取温敏核不育水稻花药总蛋白质,利用pH3-10线性胶条分离,经银染显色后检测到约1,000个     

Changes in lipids and polysaccharides during pollen ontogeny in Oenothera anthers

Summary Sudan B and PAS reactions were used for the histochemical detection of lipids and polysaccharides with light microscopy in semithin slides of resin-embedded Oenothera hookeri De Vries anthers at different ontogenic stages. The aim was to establish the variations that occur in both kinds of metabolic products during normal pollen development.In reproductive cells a wave of amylolysis/amylogenesis is observed simultaneously with a lipidic wave. Starch in diploid PMCs is different from starch in the haploid vegetative cell. The gametogenic cell produces neither reserve lipids nor starch. Callose is seen as brilliant white with Sudan B, but is PAS negative at the PMC stage, becoming PAS positive at the tetrad stage. The exine has interbedded lipopolysaccharidic materials. Tapetal plastids show an intense production of lipidic bodies after the vacuolated microspore stage. Lipidic pollen-kitt is present in the immature pollen grains and becomes dissolved at anthesis. Tapetal cells have no cellulosic cell walls from the PMC stage onwards, but they do have a glycocalix. These events have evolutionary significance and will be compared with male-sterile development.Grant of the CONICET-PID 3014000/88 and of the CONICOR-Sub. number 1574/90

---

     

Genetic control of male fertility in Arabidopsis thaliana: structural analyses of postmeiotic developmental mutants

Seven new male-sterile mutants (ms7–ms13) of Arabidopsis thaliana (L.) Heynh. (ecotype columbia) are described that show a postmeiotic defect of microspore development. In ms9 mutants, microspores recently released from the tetrad appear irregular in shape and are often without exines. The earliest evidence of abnormality in ms12 mutants is degeneration of microspores that lack normal exine sculpturing, suggesting that the MS12 product is important in the formation of pollen exine. Teratomes (abnormally enlarged microsporocytes) are also occasionally present and each has a poorly developed exine. In ms7 mutant plants, the tapetal cytoplasm disintegrates at the late vacuolate microspore stage, apparently causing the degeneration of microspores and pollen grains. With ms8 mutants, the exine of the microspores appears similar to that of the wild type. However, intine development appears impaired and pollen grains rupture prior to maturity. In ms11 mutants, the first detectable abnormality appears at the mid to late vacuolate stage. The absence of fluorescence in the microspores and tapetal cells after staining with 4′,6-diamidino-2-phenylindole (DAPI) and the occasional presence of teratomes indicate degradation of DNA. Viable pollen from ms10 mutant plants is dehisced from anthers but appears to have surface abnormalities affecting interaction with the stigma. Pollen only germinates in high-humidity conditions or during in-vitro germination experiments. Mutant plants also have bright-green stems, suggesting that ms10 belongs to the eceriferum (cer) class of mutants. However, ms10 and cer6 are non-allelic. The ms13 mutant has a similar phenotype to ms10, suggesting is also an eceriferum mutation. Each of these seven mutants had a greater number of flowers than congenic male-fertile plants. The non-allelic nature of these mutants and their different developmental end-points indicate that seven different genes important for the later stages of pollen development have been identified. Received: 14 August 1997 / Accepted: 7 October 1997     

Calcium distribution in fertile and sterile anthers of a photoperiod-sensitive genic male-sterile rice

Potassium antimonate was used to locate Ca²⁺ in fertile and sterile anthers of a photoperiod-sensitive genic male-sterile rice (Oryza sativa L. japonica). During the development of fertile anthers, abundant calcium precipitates accumulated in the anther walls and on the surface of pollen grains and Ubish bodies at the late developmental stage of the microspore, but not in the cytoplasm of pollen grains. Following the accumulation of starch grains in pollen, calcium precipitates on pollen walls diminished and increased in parenchymatous cells of the connective tissue. In sterile anthers, calcium precipitates were abundant in the middle layer and endothecium, but not in the tapetum, as was found in fertile anthers. A special cell wall was observed between the tapetum and middle layer of sterile anthers that appeared to relate to distinctive calcium accumulation patterns and poor pollen wall formation in the loculi. The formation of different patterns of antimonate-induced calcium precipitates in the anthers of photoperiod-sensitive genic male-sterile rice indicates that anomalies in the distribution of calcium accumulation correlate with the failure of pollen development and pollen abortion. Received: 30 May 1997 / Accepted: 5 July 1997     

Induced androgenesis in tomato (Lycopersicon esculentum Mill.) I. Influence of genotype on androgenetic ability

The androgenetic ability of 85 Lycopersicon esculentum Mill. genotypes was tested. Callus was induced from anthers of 53 lines and hybrids. Regeneration of plants was obtained only from calli of 15 genotypes. The data obtained clearly showed that the genotype affects induced androgenesis in tomato. The in vitro response of anthers from the cultivars Roma, Pearson, San Marzano, Por, Sar, Vigapol, Day, David and Start, containing the ms 10³⁵ gene, which is responsible for male sterility in tomato, confirm the strongly expressed dependence of both callus induction and organogenetic potential on the homozygous or heterozygous state of that gene. A protocol of callus induction, organogenesis and plant regeneration has been developed. More than 6000 regenerants have been obtained. Most of them showed different morphological alterations and variations in chromosome number (n, 2n, 4n). Some are interesting as source material for tomato breeding. Received: 17 April 1997 / Revision received: 4 August 1997 / Accepted: 15 October 1997     

橡胶树花药和小孢子发生与发育过程的观察

应用石蜡切片法．观察橡胶树的实生树和RRIM600、GT-1品系的花药壁以及小孢子的发生和发育过程,得到如下结果：1．实生树的花药壁通常由四层细胞组成,发育形式为双子叶型。药室内壁细胞在发育后期进行径向条纹加厚．至花药开裂时仍保留着原生质体。中层由一层或不规则的两层细胞组成,在小孢子单核期消失。绒毡层细胞具单棱或双核,属分泌型,至花粉发育到三细胞时消失。小孢子母细胞减数分裂为同时型。成熟花粉粒具三十细胞。精细胞椭圆形,在光镜下不能区分出细胞质鞘和核仁。所观察的实生树雄花,多数发育正常,很少有空秕的花粉。2．RRIMB00品系的花药和小孢子发生与发育和实生树相似,但至后期只有少数花粉发育正常,多数成为大小不等的败育花粉;此外也有一些败育的雄花。3．GT-1的花药在小孢子母细胞减数分裂时,绒毡层细胞的体积开始异常增大并液泡化．小孢子在四分体内解体或分离后成为空秕花粉。     

Autoclaved and filter sterilized liquid media in maize anther culture: significance of activated charcoal

Summary Medium sterilization techniques (autoclaving, filter sterilization and separate sterilization of medium components), combined with preculture exposure to activated charcoal (AC) were evaluated for effects on maize anther culture response. The addition of AC to filter sterilized medium had no effect on the number of embryo-like-structures (ES) produced. For autoclaved medium, pre-culture AC treatment resulted in a 3-fold increase in ES yield over medium lacking AC. When AC was included, autoclaved medium was more productive than filter sterilized medium. Autoclaved media without AC gave lower response than filter sterilized medium. Separate sterilization of sucrose or FeEDTA was beneficial for media autoclaved in the absence of AC. However, when all components were autoclaved together in the presence of AC, there was no advantage to separate sterilization. The maximum ES frequency (224.6 ES/100 anthers) was obtained with the genotype ETH-M 52 cultured in autoclaved medium which had been exposed to AC (5 g/L) for 96 h prior to culture initiation. It is supposed that the higher ES frequencies observed with AC-treated, autoclaved media were due to the availability of glucose and fructose following heat-induced hydrolysis of sucrose and the AC-mediated adsorption of inhibitory compounds produced during autoclaving.