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.     

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。     

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。     

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。     

植物体内游离脯氨酸的测定

游离的脯氨酸普遍存在于植物体中,正常的花粉中含量尤多,例如在一些禾本科草类的花粉中,含量可达花粉干重的1.65％,在苹果花粉中含量可达其干重的2.2％,而在不育的花粉中含量却很低。因而花粉中游离脯氨酸的含量高低被认为与植物的育性有关,有人把它作为育性的指标。另外,在干旱环境下生长的植物,体内常有游离脯氨酸的积累,故又可把它作为抗旱性的生理指标。     

花期冻害对杜仲花粉营养成分的影响

通过测定花期冻害前后杜仲花粉营养成分和总黄酮含量,研究花期冻害对杜仲花粉营养成分的影响.结果显示,花期低温对花粉蛋白质、蛋白质氨基酸、维生素含量等没有显著的影响;总黄酮含量略有增加,还原糖含量显著增加(P<0.05);但花粉游离氨基酸含量却有较明显的减少,蔗糖和粗脂肪的含量极显著的减少(P<0.01).所以,在今后的生产中应尽量采取措施预防花粉冻害.     

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。     

南北样带温带区栎属树种种子化学组成与气候因子的关系

选取南北样带温带区8个核心分布区的7种优势栎属树种作为对象,通过检测种子的化学组成,包括淀粉、蛋白质、油脂、可溶性总糖、单宁和脯氨酸,分析其化学组成与气候因子之间的关系,结果发现:在气候因子中,温度是影响种子化学组成的主要因素,日照时数次之,而降水没有明显影响.在南北样带温带区,随纬度的增加,温度降低,年日照时数增加,生长积温减少,生长季缩短,淀粉含量积累下降,并趋向于水解为可溶性糖类;同时,从南到北,为适应低温的环境条件,栎属树种通过增加种子蛋白质、可溶性总糖和脯氨酸的含量,以提高种子的抗寒性.栎属树种种子的化学组成之间存在一定的相互关系,可溶性总糖、蛋白质和脯氨酸之间呈正相关关系,它们三者与淀粉均为负相关,种子化学组成之间的这种相互关系是植物对环境适应的生态策略.     

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...     

文心兰花芽分化过程中糖类和蛋白质的组织化学定位

以文心兰‘milliongolds’不同发育阶段的花芽为试材,研究糖类和蛋白质在文心兰花芽分化过程中的变化。经石蜡切片高碘酸席夫反应法（PAS法）和汞溴酚蓝组织化学染色观察发现,糖类和蛋白质主要以颗粒状分布在各器官的原基顶端及其周围。在花序原基分化期,细胞中糖含量较少,而蛋白质含量丰富且整体分布均匀。随着花芽的继续分化,糖和蛋白质含量在花萼原基分化期最为丰富,在花瓣原基时期含量明显减少;进入合蕊柱及花粉块分化期时,糖和蛋白质在合蕊柱及花粉块着生处最为集中。在文心兰花发育过程中,对不同分化时期中花芽的糖和蛋白质含量进行测定,表明糖和蛋白质含量的变化趋势与组织化学定位相一致。     

Inhibition of RNA and protein synthesis in pollen tube development of Pinus bungeana by actinomycin D and cycloheximide

* The effects of actinomycin D and cycloheximide on RNA and protein synthesis were investigated during pollen tube development of Pinus bungeana. * RNA and protein contents, protein expression patterns, cell wall components and ultrastructural changes of pollen tubes were studied using spectrophotometry, SDS-PAGE electrophoresis, Fourier transformed infrared (FTIR) microspectroscopy and transmission electron microscopy (TEM). * Pollen grains germinated in the presence of actinomycin D, but tube elongation and RNA synthesis were inhibited. By contrast, cycloheximide inhibited pollen germination and protein synthesis, induced abnormal tube morphology, and retarded the tube growth rate. SDS-PAGE analysis showed that protein expression patterns changed distinctly, with some proteins being specific for each phase. FTIR microspectroscopy established significant changes in the chemical composition of pollen tube walls. TEM analysis revealed the inhibitors caused disintegration of organelles involved in the secretory system. * These results suggested RNA necessary for pollen germination and early tube growth were present already in the pollen grains before germination, while the initiation of germination and the maintenance of pollen tube elongation depended on continuous protein synthesis.     

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.     

Histological and Morphological Studies of Pollen Grains from Elongata, Reduced Elongata and Staminate Flowers in Carica papaya L.

The quality and quantity of pollen from three types of flowers; elongata, reduced elongata and staminate from the commercialized Thai papaya cultivar ‘Khak Nual’ were determined using pollen morphology, pollen physical characters and pollen development processes. Pollen development progressed at the same pace in the three types of pollen-producing flowers and was consistent with pollen development in many angiosperms. Pollen morphology showed that papaya pollen grains are tricolporate, with three apertures, and there is no significant difference in diameter (25.18–25.72 μm) and weight (11.76–15.45 ng) among pollen sources. The staminate flower shows the lowest amount of pollen, with 12,368 pollen per anther, but higher viability and germination rates of 95.53 % and 53.64 %, respectively. In contrast, the elongata type shows the highest amount of pollen grains with 14,884 pollen per anther and the lowest viability and germination rates, 93.06 % and 46.33 % respectively. The physical characteristics of pollen grains from reduced elongate and elongate flowers are similar. Reduced elongate flower type can donate pollen without self-pollinating.     

桃花粉离体萌发和花粉管生长特性研究

采用花粉离体萌发法研究不同培养基组分和培养条件对桃花粉萌发和花粉管生长的影响,同时对不同贮藏温度下的桃花粉寿命进行研究.结果表明:固体培养基与液体培养基对桃的花粉萌发率和花粉管长度影响差异不显著;10%蔗糖是大多数桃品种花粉的最适萌发条件;硼能提高桃花粉的萌发率,但对花粉管的生长没有促进作用;桃花粉在20℃～25℃的培养温度下萌发率最高,花粉管最长;桃花粉萌发率和花粉管长度在培养前3 h内上升最快,3～5 h上升趋势减弱,5 h后基本停止;随着贮藏温度的升高和贮藏时间的延长,花粉生活力呈降低的趋势.     

山丹不同居群花粉形态研究

为掌握山丹居群在孢粉学方面的遗传多样性,在扫描电子显微镜下对25个居群的山丹花粉进行了形态观察,结果表明:山丹花粉粒形态为单花粉粒,极面观为椭圆体,具单萌发沟。不同居群的花粉粒在大小、网眼直径、萌发沟宽、P/E值和外壁纹饰等方面存在不同程度的差异,且居群间花粉性状的变异大于居群内的变异,最大变异系数达25.52%。     

Honey bee‐collected pollen in agro‐ecosystems reveals diet diversity,diet quality,and pesticide exposure

European honey bees Apis mellifera are important commercial pollinators that have suffered greater than normal overwintering losses since 2007 in North America and Europe. Contributing factors likely include a combination of parasites, pesticides, and poor nutrition. We examined diet diversity, diet nutritional quality, and pesticides in honey bee‐collected pollen from commercial colonies in the Canadian Maritime Provinces in spring and summer 2011. We sampled pollen collected by honey bees at colonies in four site types: apple orchards, blueberry fields, cranberry bogs, and fallow fields. Proportion of honey bee‐collected pollen from crop versus noncrop flowers was high in apple, very low in blueberry, and low in cranberry sites. Pollen nutritional value tended to be relatively good from apple and cranberry sites and poor from blueberry and fallow sites. Floral surveys ranked, from highest to lowest in diversity, fallow, cranberry, apple, and blueberry sites. Pesticide diversity in honey bee‐collected pollen was high from apple and blueberry sites and low from cranberry and fallow sites. Four different neonicotinoid pesticides were detected, but neither these nor any other pesticides were at or above LD₅₀ levels. Pollen hazard quotients were highest in apple and blueberry sites and lowest in fallow sites. Pollen hazard quotients were also negatively correlated with the number of flower taxa detected in surveys. Results reveal differences among site types in diet diversity, diet quality, and pesticide exposure that are informative for improving honey bee and land agro‐ecosystem management.     

桔梗雄性不育花药的发育解剖学研究

通过田间观察和石蜡切片法对桔梗不育和可育花药的形态及其解剖结构进行了比较研究,结果表明:桔梗可育和不育花器形态、花药长度无明显差异,而开花后,可育花药饱满、有光泽、有花粉粒散出,不育花药皱瘪、无光泽、无花粉粒散出;不育花药绒毡层提前退化解体,导致小孢子发育异常。     

In vitro pollen functionality of attacin-transgenic “Royal Gala” apple plants and apples transformed with 1-aminocyclopropane-1-carboxylic acid synthase (ACS)-antisense vector

Abstract

To assess pollen functionality of transgenic apple trees, in vitro pollen germination and tube growth were evaluated. Flowers of transgenic “Royal Gala” apple lines containing attacin E gene to confer resistance to fire blight (Erwinia amylovora), or antisense 1-aminocyclopropane-1-carboxylic acid synthase (ACS) construct to improve fruit storage life, were collected, and pollen was harvested. Amongst the 19 transgenic lines, pollen from three lines transformed with an ACS-antisense vector consistently had significantly lower germination rate compared to “Royal Gala”; however, no correlation between ACS level in fruit and pollen germination rate was observed. Western blots showed that the amounts of the lytic protein, attacin, varied in pollen of the four attacin-transgenic lines sampled. There was no significant correlation between attacin level in the pollen and pollen germination rate or pollen tube growth. The addition of boric acid to the germination buffer enhanced germination in attacin-transgenic lines, as well as in lines down-regulated in ethylene synthesis and control “Royal Gala”. This initial study suggests that the majority of transgenic lines tested do not differ from the control “Royal Gala” in pollen germination, and that attacin or down-regulation of ethylene does not influence in vitro pollen functionality.     

基于响应面法的白蜡属花粉离体萌发培养基优化

基于Box-Behnken设计(Box-Behnken design, BBD)的响应面法, 对绒毛白蜡(Fraxinus velutina)、新疆小叶白蜡(F. sogdiana)和美国白蜡(F. americana)3个树种花粉的离体萌发培养基成分进行了研究。以花粉萌发率为响应指标, 建立了3种培养基成分(蔗糖、CaCl₂和H₃BO₃)与花粉萌发率间的响应关系模型。在此基础上, 通过无约束优化设计得到了3个树种花粉的最佳萌发条件。 结果表明, 蔗糖是影响花粉萌发的最主要因素, 当蔗糖浓度一定时, CaCl₂和H₃BO₃之间交互作用明显。同时还对响应面建模优化后得到的最佳萌发培养基进行了验证, 结果表明: 上述3种花粉的实际萌发率依次为58.33%、69.71%和59.42%, 均与优化得到的理论响应值相吻合, 同时也验证了基于BBD响应面模型进行花粉离体萌发条件优化方法的有效性。     

The block of intracellular calcium release affects the pollen tube development of Picea wilsonii by changing the deposition of cell wall components

Two potent drugs, neomycin and TMB-8, which can block intracellular calcium release, were used to investigate their influence on pollen tube growth and cell wall deposition in Picea wilsonii. Apart from inhibiting pollen germination and pollen tube growth, the two drugs largely influenced tube morphology. The drugs not only obviously disturbed the generation and maintenance of the tip-localized Ca(2+) gradient but also led to a heavy accumulation of callose at the tip region of P. wilsonii pollen tubes. Fourier transform infrared (FTIR) spectroscopy analysis showed that the deposition of cell wall components, such as carboxylic acid, pectins, and other polysaccharides, in pollen tubes was changed by the two drugs. The results obtained from immunolabeling with different pectin and arabinogalactan protein antibodies agreed well with the FTIR results and further demonstrated that the generation and maintenance of the gradient of cross-linked pectins, as well as the proportional distribution of arabinogalactan proteins in tube cell walls, are essential for pollen tube growth. These results strongly suggest that intracellular calcium release mediates the processes of pollen germination and pollen tube growth in P. wilsonii and its inhibition can lead to abnormal growth by disturbing the deposition of cell wall components in pollen tube tips.