峨眉拟单性木兰雄性不育的细胞形态学及生理生化特性研究

峨眉拟单性木兰(Parakmeria omeiensis)是木兰科(Magnoliaceae)拟单性木兰属(Parakmeria)的常绿乔木,属于国家Ⅰ级保护的极度濒危植物。为探究峨眉拟单性木兰两性花中雄性败育发生的时期及花药不同发育时期的生理生化特性,以两性花中的不育雄蕊和雄花中的可育雄蕊为材料,利用石蜡切片观察2种雄蕊的花药发育过程,并测定不同发育时期的可溶性糖、可溶性蛋白、脯氨酸的含量,分析过氧化氢酶(CAT)、过氧化物酶(POD)的活性。结果表明：(1)不育雄蕊与可育雄蕊在减速分裂时期出现明显差异,不育雄蕊的绒毡层致密、没有发育,四分体未形成,随后解体,花粉囊中无花粉;可育雄蕊的绒毡层和小孢子母细胞发育正常,成熟时花粉囊开裂,花粉粒溢出。(2)不育雄蕊的可溶性糖含量、可溶性蛋白含量、脯氨酸含量,在减数分裂时期、单核期和花粉成熟期都显著低于可育雄蕊。(3)不育雄蕊POD活性整体呈上升趋势,在减数分裂时期、单核期和花粉成熟期都显著高于可育雄蕊;不育雄蕊CAT活性整体呈下降趋势,显著低于同时期的可育雄蕊。综上认为,两性花中雄性败育发生在减速分裂时期,其败育的主要原因是物质能量代谢降低...     

百合育性与花药发育过程中激素和内源物质含量及能量代谢酶活性的关系

为揭示百合雄性不育发生与其花药内源激素含量、几种内源物质含量和能量代谢酶活性变化的关系,该研究利用高效液相色谱法、串联质谱法和比色法,测定了可育系和不育系花药发育关键时期的激素、可溶性糖、可溶性蛋白、丙二醛、脯氨酸含量和COD及ATP能量代谢酶活性,分析激素含量、各激素间比值、内源物质含量和能量代谢酶活性对百合雄性不育的影响。结果表明:(1)在百合花药造孢细胞期到四分体期,花药内源激素含量、内源物质含量及能量代谢酶活性变化在不育系与可育系间均存在差异。其中,不育系花药的生长素(IAA)含量在发育的各阶段均高于可育系;ZR和GA4含量在花粉母细胞期不育系与可育系相差不大,但到四分体期不育系中的含量低于可育系;ABA含量在不育系四分体期出现盈积;JA含量在不育系的花粉母细胞期低于可育系,而在四分体期高于可育系。(2)花药内源物质中可溶性糖、可溶性蛋白和脯氨酸含量表现为可育系均高于不育系,但不育系的丙二醛含量在四分体期显著高于可育系。(3)COD和ATP能量代谢酶活性在花药发育的各阶段均是可育系高于不育系。研究认为,在百合不育系花药发育关键时期(花粉母细胞期到四分体期),其IAA含量增加,ABA水平提高,GA4和ZR含量降低,影响了花药内源物质和能量的正常代谢,这可能是导致百合花药中花粉发育受阻、形成败育的原因。     

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。     

枸杞花粉发育进程中花蕾和叶片物质代谢与育性的关系

以宁夏枸杞主栽品种'宁杞1号'的雄性不育株和可育株为研究材料,通过测定它们枝条生长速率,花粉不同发育时期花蕾和叶片的叶绿素、可溶性糖和脯氨酸含量以及硝酸还原酶活性,比较不育株与可育株在发育进程中物质代谢的差异.结果表明:在枸杞花粉发育进程中,不育株枝条生长速率快,花蕾和叶片的叶绿素含量高,而脯氨酸严重缺乏,可溶性糖含量和硝酸还原酶活性低,且各个发育阶段不育株和可育株均存在明显差异.可见,'宁杞1号'不育株物质代谢水平低,缺乏生理活性物质积累,使花粉发育有关基因的表达受到抑制,最终影响了其育性的正常表达.     

红麻细胞质雄性不育系与保持系花药活性氧代谢差异比较

以红麻细胞质雄性不育系L23A及其保持系L23B为材料,比较其花药淀粉及可溶性糖含量变化并分析呼吸速率、活性氧产生速率、丙二醛(MDA)含量以及活性氧清除酶(POD、SOD)含量变化,来探讨活性氧伤害与红麻雄性不育的关系。结果表明:在小孢子发育的单核期,不育系呼吸速率与保持系差异不明显,但不育系花药O^-₂·含量高于保持系; 在双核期,不育系的呼吸速率明显低于其保持系,但不育系花药O^-₂·含量与保持系花药相近; 不育系在单核期和双核期的呼吸速率几乎没有变化,而保持系同一时期的呼吸速率呈明显增高趋势; 在不育系败育过程中,药隔维管组织中的大颗粒淀粉含量几乎不变,且不育系花药中的可溶性糖含量在单核期和双核期均低于保持系。推测是由于不育系花药中抗氰呼吸降低,一方面导致花药物质代谢和能量代谢的紊乱,不育系花药不能利用药隔组织中的淀粉粒,另一方面不能有效将细胞内过多电子通过抗氰呼吸传至O₂,引致不育花药中O^-₂·升高,从而导致MDA含量在单核期和双核期均高于保持系,同时POD的活性在单核期及双核期均低于保持系,而SOD活性在单核期高于保持系,在双核期则低于保持系。不育系花药在发育中,花药O^-₂·和MDA过量积累,以及SOD和POD酶活性降低,导致活性氧产生与清除失去平衡,花粉败育。     

化学杂交剂诱导的小麦生理型雄性不育花药的活性氧代谢

以化学杂交剂SQ-1诱导的生理型小麦雄性不育及其对照植株花药为材料,研究了不同花粉发育时期花药中超氧阴离子(O-·2)生成速率、过氧化氢(H2O2)和丙二醛(MDA)含量以及主要抗氧化酶活性的变化,以探明小麦花药活性氧代谢和生理型雄性不育的关系.结果表明,在幼穗时期,O-·2生成速率、H2O2和MDA含量、超氧化物歧化酶(SOD)和抗坏血酸过氧化物酶(APX)活性均高于相应对照,而过氧化物酶(POD)和过氧化氢酶(CAT)活性则低于或显著低于对照;在单核早期以及花粉败育主要发生期(单核后期和二核初期),O-·2生成速率、H2O2和MDA含量极显著高于对照,而SOD、POD、CAT和APX酶活性却极显著低于对照;在败育后的花药中,O-·2生成速率和H2O2含量与对照之间差异幅度缩小,但MDA含量依然加大,同期的几种抗氧化酶活性依然极显著低于对照.在败育的关键期,品种'西农1376'处理株花药的活性氧升高幅度比'西农2611'处理株较大,抗氧化酶活性降低幅度也较大,且'西农1376'处理株的相对雄性不育率也较高.可见,化学杂交剂SQ-1能诱导小麦花药中O-·2和H2O2大量积累以及SOD、POD、CAT和APX活性的极显著降低,引起花粉关键败育期花药活性氧代谢严重失衡和严重膜脂过氧化,导致大量花粉母细胞发育受到严重抑制,最终造成小麦生理型雄性不育.     

高羊茅花粉母细胞减数分裂及其雄性不育的细胞生理学研究

为探讨高羊茅雄性不育株A22013189的小孢子发育过程及其败育的生理学机理,以可育株189为对照,对其结实能力、花粉活力、花粉母细胞减数分裂进程中的染色体行为及生理生化特征进行研究。结果表明:(1)不育株A22013189花粉数量少,花粉粒空瘪皱褶,自交不结实,在杂交中作父本获得杂交后代的可能性极小。(2)从减数分裂前期Ⅰ至四分体时期,不育株A22013189花粉母细胞存在大量落后染色体、染色体桥和断片、微核、单价染色体、不均等分离、染色体分裂不同步、游离染色体、三分体、60°纺锤体、染色体缺失等异常现象,初步分析这些小孢子异常分裂是导致高羊茅花粉败育的细胞学原因之一。(3)不育株A22013189的可溶性蛋白质、可溶性糖含量在整个发育时期都显著低于同期可育株189;苗期至造孢细胞期,A22013189游离脯氨酸含量与可育株189无显著差异,但减数分裂期至花粉成熟期,A22013189游离脯氨酸含量却显著低于同期可育株189;苗期至造孢细胞期,A22013189丙二醛含量显著低于同期可育株189,但小孢子进入减数分裂期后,A22013189丙二醛的增加速度和积累量明显高于同期可育株189。研究发现,高羊茅雄性不育株花粉母细胞减数分裂染色体行为异常,生长发育过程存在物质能量代谢降低,有害物质积累现象。研究结果对于高羊茅败育机理研究及杂交育种亲本选择有重要的理论指导意义。     

苎麻雄性不育材料的生理生化特性初探

在苎麻(Boehmeria nivea(L.) Gaud)雌雄性器官发育期取功能叶,现蕾后取幼蕾(直径小于0.4mm)、中蕾(直径0.4-1.2mm)、大蕾(直径大于2.0mm),比较分析苎麻雄性不育材料C26、C4与可育材料B8、B16的可溶性糖含量、淀粉含量、游离脯氨酸含量、可溶性蛋白含量以及过氧化物酶(POD)活性的差异。结果表明,不育材料在各时期的花蕾中可溶性糖含量都高于可育材料的,以大蕾最为明显,C26比B8高0.58%,C4比B16高0.83%;淀粉和游离脯氨酸含量从中蕾期开始,不育材料明显低于可育材料,到大蕾期差异最大(淀粉含量B8/C26为1.81,B16/C4为1.46;游离脯氨酸含量B8是C26的7.35倍,B16为C4的3.94倍);在相同花蕾期,可育材料的POD活性略高于不育材料。可溶性蛋白质含量在可育材料和不育材料中无明显规律性。这些物质的代谢可能是导致苎麻雄性不育的原因。     

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。     

葱胞质雄性不育花蕾生化物质含量和能量代谢酶活性的动态变化特征

以葱胞质雄性不育系CA及其同核异质保持系CB为试材,研究了花蕾发育过程中IAA、GA3、ZR、ABA含量以及细胞色素氧化酶(COD)和ATP酶(ATPase)活性、可溶性糖、游离氨基酸和可溶性蛋白质含量的动态变化规律.结果显示:(1)葱不育系花蕾的IAA和GA3含量在败育过程中显著低于保持系,而ZR和ABA含量则较保持系有不同程度的盈积.不育系花蕾COD和ATPase活性以及游离氨基酸含量、可溶性蛋白质含量在中蕾期后显著低于保持系,可溶性糖在不同发育时期均基本低于保持系.(2)不育系花蕾IAA和GA3含量与可溶性糖含量变化均呈显著正相关,不育系ZR和保持系ABA含量则分别与COD、ATPase活性以及可溶性糖、可溶性蛋白质含量存在负相关关系.研究表明,葱胞质雄性不育系花蕾中IAA和GA3亏缺,而ZR和ABA盈余,各种内源激素含量和营养物质含量与能量代谢有关酶活性相关性不尽相同.     

Anther starch variations in Lilium during pollen development

Starch was cytologically localized and biochemically assayed in different anther cell layers of Lilium cv. Enchantment during pollen development and its presence was correlated with anther growth. Two phases could be distinguished: the first, the growth phase, extends from the beginning of meiosis to the vacuolated microspore stage and corresponds to maximum increase in anther size and weight. During this period, microspores lack amyloplasts and starch is degraded in the outer staminal wall layers. The tapetum does not contain starch reserves but accumulates a PAS-positive substance in its vacuole. The second phase, the maturation phase, begins with the late vacuolated microspore stage and lasts until pollen maturation. Anther growth is slowed during this phase. A wave of amylogenesis/ amylolysis occurs first in the late vacuolated-microspores and young pollen grains and, next, in the staminal envelopes. In the pollen grain, the cytoplasm of the vegetative cell is filled with starch, but amyloplasts are not detected in the generative cell. When pollen grains ripen, amylaceous reserves are replaced with lipids. In the staminal envelopes, the second amylogenesis is particularly evident in the endothecium and the middle layers; the peak of starch is reached at the young bicellular pollen grain stage; starch disappears from the anther wall early during the maturation phase. The wave of amylogenesis/amylolysis occurring in the staminal envelopes during the maturation phase is peculiar to Lilium. It is interpreted as a sudden increase in carbohydrate level caused by lower anther needs when the growth is completed. Staminal envelopes may act as a physiological buffer and regulate soluble sugar level in the anther. Stages of anther growth correlate with starch content variations and this suggests that during the growth phase, products of starch hydrolysis in the staminal envelopes may be consumed partly by anther cell layers and partly by microspores.     

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.     

凤仙花花药发育中多糖和脂滴组织化学研究

以不同发育时期的凤仙花花药为实验材料,采用组织化学方法,对花药发育中的结构变化及多糖和脂滴物质分布进行观察。结果表明:(1)凤仙花的花药壁由6层细胞组成,包括1层表皮细胞,2层药室内壁细胞,2层中层细胞和1层绒毡层细胞。其中绒毡层细胞的形态不明显,很难与造孢细胞区分,且在小孢子母细胞时期退化。(2)在小孢子母细胞中出现了一些淀粉粒,但减数分裂后,早期小孢子中的淀粉粒消失,又出现了一些小的脂滴;随着花粉的发育,小孢子形成大液泡,晚期小孢子中的脂滴也消失;小孢子分裂形成二胞花粉后,营养细胞中的大液泡降解、消失,二胞花粉中又开始积累淀粉;接近开花时,成熟花粉中充满细胞质,其中包含了较多的淀粉粒和脂滴。(3)在凤仙花的花药发育中,绒毡层细胞很早退化,为小孢子母细胞和四分体小孢子提供了营养物质;其后的中层细胞退化则为后期花粉发育提供了营养物质。     

Assessing hybrid sterility in <Emphasis Type="Italic">Oryza glaberrima</Emphasis> × <Emphasis Type="Italic">O. sativa</Emphasis> hybrid progenies by PCR marker analysis and crossing with wide compatibility varieties

Interspecific crossing of the African indigenous rice Oryza glaberrima with Oryza sativa cultivars is hindered by crossing barriers causing 100% spikelet sterility in F₁ hybrids. Since hybrids are partially female fertile, fertility can be restored by back crossing (BC) to a recurrent male parent. Distinct genetic models on spikelet sterility have been developed predicting, e.g., the existence of a gamete eliminator and/or a pollen killer. Linkage of sterility to the waxy starch synthase gene and the chromogen gene C, both located on chromosome 6, have been demonstrated. We selected a segregating BC₂F₃ population of semi-sterile O. glaberrima × O. sativa indica hybrid progenies for analyses with PCR markers located at the respective chromosome-6 region. These analyses revealed that semi-sterile plants were heterozygous for a marker (OSR25) located in the waxy promoter, whereas fertile progenies were homozygous for the O. glaberrima allele. Adjacent markers showed no linkage to spikelet sterility. Semi-sterility of hybrid progenies was maintained at least until the F₄ progeny generation, suggesting the existence of a pollen killer in this plant material. Monitoring of reproductive plant development showed that spikelet sterility was at least partially due to an arrest of pollen development at the microspore stage. In order to address the question whether genes responsible for F₁ sterility in intraspecific hybrids (O. sativa indica × japonica) also cause spikelet sterility in interspecific hybrids, crossings with wide compatibility varieties (WCV) were performed. WCV accessions possess "neutral" S-loci (Sⁿ) improving fertility in intraspecific hybrids. This experiment showed that the tested Sⁿ-loci had no fertility restoring effect in F₁ interspecific hybrids. Pollen development was completely arrested at the microspore stage and grains were never obtained after selfing. This suggests that distinct or additional S-loci are responsible for sterility of O. glaberrima × O. sativa hybrids.Communicated by H.C. Becker     

车桑子小孢子发生与雄配子体发育

为了解干热河谷区车桑子(Dodonaea viscosa)胚胎学特征及其结籽率低的原因,采用常规石蜡切片法和电镜扫描技术对车桑子小孢子发生、雄配子体发育和花粉的形态特征进行了观察。结果表明,车桑子花药具有4个花粉囊。完整的花药壁从外到内依次为表皮、药室内壁、2～3层中层细胞和绒毡层;绒毡层类型是腺质绒毡层。花药成熟期,中层、绒毡层均退化消失。小孢子母细胞进行同时型胞质分裂;四分体为四面体型结构。成熟的花粉为二细胞型。花粉近球形,外壁密布颗粒状纹饰,具有3条不构成合沟的萌发沟。雄性生殖发育过程出现的异常现象可能是干热河谷地区车桑子结籽率低的原因之一。     

Sucrose and starch catabolism in the anther of <Emphasis Type="Italic">Lilium</Emphasis> during its development: a comparative study among the anther wall,locular fluid and microspore/pollen fractions

In order to better understand the various pathways of sucrose and starch catabolism in the anther of lily (Lilium hybrida var. “Enchantment”), invertase (EC 3.2.1.26) and amylase (EC 3.2.1.1, EC 3.2.1.2) activities were measured separately in different fractions (anther wall, locular fluid and microspore/pollen) and correlated with the sugar content during anther development. Our findings showed significant differences among the fractions analyzed, suggesting that the regulation of sucrose and starch catabolism could follow distinct pathways in each fraction. Glucose and fructose amounts progressively decreased from anther wall to fluid and from fluid to microspore/pollen. Thus, the developing pollen could act as a sink for the carbohydrates that reach the anther. In this sense, cell wall-bound invertases seem to play a major role in soluble sugar partitioning in the different fractions of the anther. Sucrose concentration was found to be substantially higher in the locular fluid than in the other fractions, indicating a probable site for storage. On the other hand, the anther wall tissues could have a buffering function, storing nutrient surplus in starch grains and thus regulating the availability of soluble sugars in the whole anther. All these results proved the advantages of the experimental model proposed here, as well as its usefulness to investigate sugar metabolism in Lilium anthers.     

长花柱型滇丁香小孢子发生及雄配子体发育

利用常规石蜡切片法和细胞学压片法,对异型花柱植物滇丁香的长花柱型植株的小孢子发生、雄配子体发育及花粉萌发进行观察。结果表明:(1)长花柱型滇丁香具5枚花药,花药4室。(2)花药壁由1层表皮、1层花药内壁、2层中层和1~3层绒毡层组成;花药壁发育方式为基本型,绒毡层类型为腺质绒毡层。(3)小孢子母细胞减数分裂的胞质分裂为同时型,四分体排列方式为四面体型,偶有左右对称型;不同药室间小孢子母细胞减数分裂不同步。(4)成熟花粉粒为二细胞型。(5)小孢子发生和雄配子体发育过程正常,表明长花柱型滇丁香属于发育正常的两性花。(6)授粉4h后,长花柱型滇丁香的花粉在长、短两种花柱柱头上的萌发率分别达(91.8±1.6)%和(93.2±1.1)%,且两者间无显著性差异(t=1.585,df=8,p=0.152),表明长花柱型滇丁香的成熟花粉粒在长、短两种花柱柱头上的萌发均正常。     

灯盏花雄性不育种质鉴定与花药发育的细胞学研究

对云南泸西栽培灯盏花群体进行调查,发现了灯盏花雄性不育种质个体,其出现频率约为1.06×10-4.对所发现的灯盏花不育株形态特征及其花药发育过程进行了观察,并对花粉活力进行鉴定.结果显示:(1)灯盏花不育株根、茎、叶形态与正常可育植株基本相似,管状花小,花丝短,花药瘦小,无花粉粒散出或花粉无活力.(2)灯盏花在其花药发育的小孢子母细胞时期、四分体时期、小孢子时期和单核早期,由于绒毡层细胞液泡化、提前解体,不能为小孢子或花粉发育提供所需物质,导致小孢子母细胞和四分体解体,产生无花粉的花药;或小孢子和单核花粉胞内降解,形成不同形状和外壁纹饰的败育花粉.研究认为,灯盏花花药绒毡层异常是其花粉败育的主要原因.     

Antisense inhibition of a nuclear gene,BrDAD1, in Brassica causes male sterility that is restorable with jasmonic acid treatment

Male sterility is widely used for the production of hybrid seeds, but the use of genic male sterility is rather limited because of difficulty in maintaining homozygous male sterile plants. Recently, the DEFECTIVE IN ANTHER DEHISCENCE 1 (DAD1) gene, which encodes a phospholipase A1 involved in the first step of the jasmonic acid (JA) biosynthesis pathway, was isolated from a male sterile Arabidopsis mutant. To utilize this gene in Brassica crops, we characterized the BrDAD1 gene, the putative ortholog of DAD1 in Brassica rapa. Out of 25 plants transformed with an antisense gene constructed from the BrDAD1, 3 plants showed a defect of anther dehiscence at the flower bud opening stage and produced inviable pollen. One of the three showed male sterility only, but the other two showed a delay or a lack of flower opening in addition to male sterility. The male sterile and flower-opening phenotypes were rescued by the application of JA as well as linolenic acid. Furthermore, all these characteristics were inherited to the next generation. The present results demonstrate a novel control system for hybrid seed production by the use of nuclear genes.