花序开放程度、栽培温度和贮藏条件对菊花品种‘钟山金阳’花粉萌发的影响

采用离体培养法和杂交实验法,以菊花品种‘钟山金阳’(Chrysanthemum morifolium‘Zhongshanjinyang’)为实验材料,比较了不同的花序开放程度(花序开放天数和散粉阶段)、栽培温度和贮藏条件对其花粉萌发的影响。离体培养结果表明:在花序开放的第2至第7天花粉萌发率为22.6%~17.3%,第9天为7.7%,第11天仅为2.8%;在散粉的第1阶段(第1和第2轮管状花散粉)和第2阶段(第3和第4轮管状花散粉)花粉萌发率分别为19.5%和17.8%,第3阶段(全部管状花散粉)花粉萌发率仅为9.4%,表明随花序的开放和散粉阶段的延续,花粉萌发率均呈逐渐降低的趋势。在昼/夜温度5℃/0℃、10℃/5℃、15℃/10℃、20℃/15℃、25℃/20℃和30℃/25℃条件下栽培一定时间,其花粉萌发率和花粉管长度均有明显差异;其中,10℃/5℃栽培组培养4.0 h的花粉萌发率最高(23.0%),而30℃/25℃栽培组培养0.5 h的花粉萌发率最低(1.2%);15℃/10℃和20℃/15℃栽培组培养4.0 h的花粉管长度较长(分别为144.4和146.1μm),而30℃/25℃栽培组培养0.5 h的花粉管长度最短(38.3μm)。在不同贮藏温度下[4℃、-20℃和室温(约25℃)],新鲜和干燥花粉的萌发率均随贮藏时间的延长逐渐下降,但降幅存在明显差异;其中,新鲜花粉贮藏12 d后花粉萌发率从21.4%降至5.0%以下,干燥花粉贮藏26 d后花粉萌发率降至5.0%以下;总体上看,干燥花粉的萌发率和贮藏时间均高于新鲜花粉,以-20℃条件下贮藏的干燥花粉萌发率最高且可贮藏至50 d以内。杂交实验结果显示:用不同栽培温度下的品种‘钟山金阳’的花粉对品种‘钟山粉碟’(‘Zhongshanfendie’)进行人工授粉,在柱头上花粉萌发数差异明显;其中,10℃/5℃栽培组的花粉萌发数最多(55.5),30℃/25℃栽培组的花粉萌发数最少(8.8),且二者间差异显著(P0.05)。综合分析结果表明:花序开放天数、散粉阶段、栽培温度及贮藏条件对菊花品种‘钟山金阳’的花粉萌发率均有明显影响。根据研究结果,建议菊花品种‘钟山金阳’花粉的最佳采集时期为花序开放的7 d内且为第1至第4轮管状花散粉阶段,栽培温度应维持在10℃左右,而且采集的花粉应置于-20℃干燥条件下贮藏。     

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.     

百合不同杂交系品种花粉贮藏特性分析

百合花粉贮藏是解决育种过程中花期不遇以及种质资源保存的重要环节,该试验以亚洲百合、东方百合和OT杂种系列的12个品种为材料,筛选出适合大部分品种花粉萌发的培养基及萌发率的观测时间,探讨花蕾发育时期[完全显色(-1级)、2/3显色(-2级)]、花粉收集后处理方式[离心管(处理Ⅰ)、离心管口加棉花(处理Ⅱ)、离心管口加棉花和管底添加干燥剂(处理Ⅲ)]、贮藏温度(常温、4℃、-20℃和-80℃)对花粉活力的影响。结果显示:(1)5%蔗糖+100(150)mg/L硼酸+30(50)mg/L氯化钙为适合大部分品种花粉萌发的液体培养基配方,百合花粉培养24h后测定萌发率为宜。(2)-1级花粉的萌发率比-2级的高且保持生命力的时间更长,如‘Tresor’-1级新鲜花粉的萌发率可达到83.2%,但其-2级只有21%。常温条件最不利于花粉保存,20d时大部分花粉萌发率为0;4℃与-20℃下较利于花粉短期贮藏且差别不显著。(3)经过4℃、120d贮藏,亚洲百合4个品种花粉萌发率最低能保持24.7%,最高能有55.9%左右;2个东方百合品种4℃下花粉萌发率为6.8%～12.7%;花粉在-80℃条件能较长时间保持活力,亚洲百合和东方百合花粉萌发率分别能保持26.7%～61.9%和7.1%～16.2%。(4)各个品种在不同温度条件下,大部分在处理Ⅲ下花粉萌发率较其他两种处理稍高。因此,-80℃结合在离心管口加棉花和管底添加干燥剂(处理Ⅲ)适合百合花粉的长期贮藏。     

秋水仙素诱导月季品种‘月月粉’2n花粉的研究

为获得二倍体古老月季品种‘月月粉’人工诱导2n花粉用于现代月季杂交育种,该研究在了解‘月月粉’花粉母细胞发育过程的基础上,使用秋水仙素注射结合包裹法处理幼嫩花蕾,成功获得了可正常萌发的2n花粉,并通过观察诱导获得的花粉离体及其在雌蕊上的萌发特征,初步评估2n花粉用于杂交育种的性能。结果表明:(1)秋水仙素药棉包裹造成部分花蕾在采粉前死亡,花蕾存活率随秋水仙素浓度提高和处理时间延长均极显著降低。(2)2n花粉诱导率在秋水仙素浓度为2.5、5.0、7.5和10.0 g/L间存在显著差异,但在处理时间为24、48和72 h间无显著差异;最佳诱导处理为5.0 g/L秋水仙素处理24 h,诱导率可达15.83%,该处理获得的2n花粉经亚历山大染色检验活力为27.2%。(3)‘月月粉’花粉母细胞在减数分裂时存在异常落后染色体和平行纺锤体,在四分体时期形成染色体数目加倍的二分体和三分体,最终产生2n花粉。(4)所诱导的2n花粉的体外萌发率和花粉管长度与天然花粉无显著差异,且都可以在母本柱头上萌发并长出花粉管进入子房,表明可用于进一步的杂交育种。     

紫斑牡丹单瓣与半重瓣花粉超微结构及生理生化特性比较北大核心CSCD

该研究以3个紫斑牡丹品种(‘象牙白’、‘美人面’、‘紫金冠’)的单瓣与半重瓣花粉为材料,对其超微结构和生理生化特性进行观察分析,并对其花粉培养基进行筛选优化,探讨单瓣和半重瓣花粉的超微结构与花粉萌发率以及物质代谢与花粉萌发率之间的关系。结果表明:(1)紫斑牡丹花粉萌发的最适宜培养基为100 g·L^(-1)蔗糖+0.15 g·L^(-1)硼酸+10 g·L^(-1)琼脂。(2)花粉饱满率是影响花粉萌发的主要原因。(3)3个紫斑牡丹品种的单瓣花粉离体培养萌发率均显著高于半重瓣的花粉。(4)‘象牙白’、‘美人面’、‘紫金冠’单瓣花粉可溶性蛋白含量均高于半重瓣花粉,但三者单瓣花粉的MDA含量和3种保护酶(CAT、SOD、POD)活性均低于半重瓣花粉。研究认为,花粉体内蛋白质亏损和MDA、CAT、SOD、POD的积累可能是引起半重瓣紫斑牡丹花粉萌发率较低的生理原因之一。     

不同脱水处理对桔梗花粉离体萌发及贮藏性能的影响

对桔梗花粉离体萌发条件进行了筛选,并采用花粉离体萌发法测定了不同干燥方式和贮藏时间处理对桔梗花粉生活力的影响;同时采用人工授粉的方法研究了不同处理后桔梗花粉授粉结实能力的变化。结果表明：（1）30℃培养1.5h是桔梗花粉离体萌发和花粉管生长的最佳培养条件。（2）不同干燥方式、不同干燥时间处理后桔梗花粉萌发率和花粉管长度均有所下降,但下降幅度不同,其中以干燥箱45℃烘干处理1.5h~2h下降最少,其花粉耐藏性也有明显提高。（3）不同干燥方式处理桔梗花粉后其授粉结籽率的变化明显不同,阴置2h处理的花粉结籽率最低,而干燥器内放置6h、日晒2h和烘干1.5h处理后花粉结籽率略有变化;干燥处理后随贮藏时间的增加,其授粉结籽率均呈下降趋势,但45℃烘干1.5h处理的花粉结籽率下降速度最慢,其授粉结籽能力可保持5d左右。     

梅花花粉离体萌发和花粉管生长研究

(南京农业大学园艺学院,南京210095)摘要:研究培养基成分、pH值和培养方式对梅花花粉离体萌发和花粉管生长的影响。结果表明:不同品种梅花花粉离体萌发的最适培养基为ME3+200g.L-1PEG4000(pH5.0),品种‘淡丰后’、‘久观绿萼’、‘喧妍宫粉’和‘月光玉蝶’最高萌发率可分别达到58.6%、60.6%、85.6%和50.7%。PEG4000能显著促进梅花花粉萌发,在培养基各成分中作用最大,不可替代。低浓度(50g.L-1)蔗糖对梅花品种花粉萌发作用不显著,而高浓度(≥100g.L-1)蔗糖明显抑制花粉萌发和花粉管生长。固体和液体培养对梅花花粉离体萌发的影响差异不显著。     

果梅花粉离体萌发及花粉管生长特性研究

研究了果梅(Prunus mumeSieb.1et Zucc.)花粉在不同培养基组分、花粉不同培养密度和不同温度及培养时间的离体萌发和花粉管生长特性.结果表明:细叶青花粉萌发及花粉管生长最适宜的液体培养基为30mmol/L MES(pH 6.5)缓冲液中含20%蔗糖,0.01%硼酸,20%PEG-4000,0.03?(NO3)2?4H2O,0.02%Mg-SO4?7H2O;萌发率达45.03%,花粉管长度达597.2μm.适宜于果梅花粉萌发和花粉管生长的花粉粒密度为20~80粒/μL.培养温度过高或过低都不利于果梅花粉的萌发和生长,25℃时花粉萌发和花粉管生长最好.细叶青"、月世界"、莺宿"3个品种的平均萌发率为48.6%,平均花粉管长度为762.3μm.果梅花粉在不同培养温度下,萌发及生长不同,在25℃条件下花粉管生长速度最快,集中在0~12 h内,3个品种花粉管平均生长速度为58.5μm/h.     

紫荆花粉离体萌发条件的研究

以紫荆(Cercis chinensis Bunge)花粉为材料,采用人工培养法研究了紫荆花粉离体萌发的最适条件.结果表明:蔗糖、硼酸在一定质量浓度范围内能促进花粉的萌发,最适花粉萌发的质量浓度分别为蔗糖50 mg/L以及硼酸20 mg/L,最佳萌发温度20℃.正交试验设计结果表明:花粉萌发的最佳质量浓度分剐为70 mg/L蔗糖以及18mg/L硼酸,最佳萌发温度为22℃,此时的萌发率可达60.51％.     

曼地亚红豆杉‘Hicksii’花粉活力检测条件优化和适宜诸藏温度分析

采用单因素实验设计对适宜于曼地亚红豆杉‘Hicksii’(Taxus media‘Hicksii')花粉活力检测的TTC染色法和离体培养法的实验条件进行了选择,并采用优化条件研究了在25℃～-196℃条件下储藏13周花粉活力的变化.结果表明:采用TTC染色法测定的花粉活力均高于离体培养法.在TTC染色法的3个影响因素(染液pH、TTC浓度和染色温度)中,染液pH对检测结果有极显著影响,而染色温度和TTC浓度则无显著影响,但温度对染色速率有影响.在离体培养法的3个影响因素(培养基中蔗糖添加量、H3BO3和Ca(NO3)2浓度及培养温度)中,蔗糖添加量对检测结果有极显著影响,在含质量体积分数15％蔗糖的培养基上花粉活力最高,而在含质量体积分数20％和25％蔗糖的培养基上花粉均不能萌发;在含100和200 mg ·L-1H3BO3的培养基中添加200 mg·L-1Ca(NO3)2均能显著提高花粉活力;培养温度对花粉萌发速率有影响但对花粉活力没有明显影响.TTC染色法的最优检测条件为:用5.0g·L-1TTC染液(pH 7.0)在35℃下染色2.0h;离体培养法的最优检测条件为:用含质量体积分数15％蔗糖、100 mg·L-1H3BO3和200 mg·L-1Ca(NO3)2的培养基暗培养4d.在25℃、4℃、0℃、-20℃、-80℃和-196℃条件下储藏13周,‘Hicksii'花粉活力和保持时间有明显差异,其中,于-80℃和-196℃储藏3d 花粉就丧失活力;于25℃和-20℃储藏7周、4 ℃储藏10周,花粉仍有一定的活力;而在0℃条件下花粉活力最高,且储藏11周花粉仍有活力.推测曼地亚红豆杉‘Hicksii’花粉对低温的抗性较差,0℃为其适宜的储藏温度.     

生态因子对滇重楼花药开裂的影响

滇重楼为延龄草科重楼属植物,具有极高的药用价值,由于重楼传统药用部位生长缓慢、繁殖力低下,以及人们对野生重楼资源的过度采挖使其资源日趋枯竭。滇重楼的花药在整个花期中存在开裂－关闭的现象,花药的有效闭合应是保护花粉、延长花粉寿命、增强雄性适合度的一种适应机制。该研究以滇重楼为对象,通过设计正交实验和对比实验,观测其花药开裂过程中的光照强度、温度、湿度的变化,探究光照强度、温度、湿度等生态因子对滇重楼花药开裂的影响以及滇重楼花药开裂与生态因子变化的关系。结果表明：(1)在滇重楼花药开裂的过程中,光照强度增强、温度升高、相对湿度下降;(2)温度是影响滇重楼花药开裂时间的主导因子,升温促进花药开裂,降温促进花药关闭;(3)高湿度及黑暗推迟花药开裂,但并不能阻止花药开裂;(4)低温可使滇重楼花药持续关闭,而光照强度越高,花药持续关闭所需的温度越低。该研究有利于解释滇重楼花药白天开裂夜晚关闭的现象与环境因子的关系,对滇重楼的栽培育种提供理论指导。     

京白梨花药开裂异常的解剖学研究

比较研究了京白梨(Pyrus ussuriensis Maxim.)、鸭梨、雪花梨花药开裂特性、环境条件对花药开裂的影响以及花药开裂异常的解剖学结构,结果表明:(1)京白梨花药开裂率明显低于鸭梨和雪花梨,使其产生花粉的量较少,从而导致传粉能力较弱.(2)京白梨、鸭梨、雪花梨在环境温度为25℃,相对湿度在 60%～80%时最适宜花药开裂,但京白梨花药开裂率始终低于鸭梨和雪花梨.(3)在京白梨花药发育后期不能形成开裂腔是引起花药不开裂的主要原因,而导致开裂腔不能形成的原因可能与单核期的绒毡层细胞不退化和药室内壁细胞不进行'U'字型加厚有关.     

高州普通野生稻主要生殖性状的数理统计分析

高州野生稻各采样点的生殖特性表现出明显的多样性。对141份野生稻的花粉育性、裂药指数、胚囊育性和结实率进行通径分析。结果表明,花粉育性、裂药指数和胚囊育性决定了结实率变异的61．17％,其中裂药指数对结实率的直接作用最大,通径系数为0．6495;花粉育性和胚囊育性对结实率的直接作用较小,通径系数分别为0．2356和0．2137,花粉育性通过裂药指数还有较大的间接通径系数（0．2960）。通过系统聚类,将高州普通野生稻分为4群,并时各群进行方差分析和邓肯氏多重比较,可归纳为：Ⅰ群的花粉育性、胚囊育性和裂药指数对结实率影响小;Ⅱ群主要由胚囊育性影响结实率;Ⅲ群主要由裂药指数影响结实率;Ⅳ群主要由花粉育性和裂药指数共同影响结实率。     

Aquaporins of the PIP2 class are required for efficient anther dehiscence in tobacco

Several processes during sexual reproduction in higher plants involve the movement of water between cells or tissues. Before flower anthesis, anther and pollen dehydration takes place before the release of mature pollen at dehiscence. Aquaporins represent a class of proteins that mediates the movement of water over cellular membranes. Aquaporins of the plasmamembrane PIP2 family are expressed in tobacco (Nicotiana tabacum) anthers and may therefore be involved in the movement of water in this organ. To gain more insight into the role these proteins may play in this process, we have analyzed their localization using immunolocalizations and generated plants displaying RNA interference of PIP2 aquaporins. Our results indicate that PIP2 protein expression is modulated during anther development. Furthermore, in tobacco PIP2 RNA interference plants, anther dehydration was slower, and dehiscence occurred later when compared with control plants. Together, our results suggest that aquaporins of the PIP2 class are required for efficient anther dehydration prior to dehiscence.     

Hermodactylus tuberosus L. (Iridaceae) pollen organisation before and after anther dehiscence

ABSTRACT

The morphology, cytology and viability of Hermodactylus tuberosus L. (Iridaceae) pollen were examined from the first mitosis until maturation and after anther opening. During maturation, the pollen coat becomes modified, and the vegetative cell cytoplasm accumulates several types of reserve substances. In the vegetative cell cytoplasm, starch is quickly utilised whereas lipid inclusions of different dimensions, shape and composition occur during pollen maturation. Pollen from opened anthers have a thin pollen coat; the cytoplasm has mostly lipid reserves, and many small vesicles and vacuoles. It is similar in size or larger than pollen located inside the anther, and its viability does not decrease until one day after anther dehiscence. Large osmiophilic bodies, different from those of the vegetative cell cytoplasm, are present in the generative cell cytoplasm starting from the first stage of pollen development. The poorly developed pollen coat in pollen from opened anthers suggests that it plays a minor role in attracting insects for pollination. The size and structural and ultrastructural features of mature pollen indicate that it does not undergo dehydration and possesses sufficient vigour for immediate germination.     

广东高州普通野生稻生殖特性的研究Ⅰ.结实率、花粉育性及其发育特点

对采自6个采样点141个编号的高州普通野生稻的结实率、花粉育性及其发育特点进行研究。结果发现,高州野生稻的结实率平均为57．23％,普遍偏低;花粉育性平均为89．3％。育性在50％以上的编号134个,占95．04％;7个编号出现不同程度的败育现象,败育类型以典败为主。其中育性最低的是GZW122,育性仅为8％;17个编号（占12,06％）的裂药性存在严重的异常现象。相关分析表明,花粉育性和裂药性与结实率存在极显著相关。不同采样点问存在一定差异,其中位于祥山镇祥山村祥山洞的编号结实率、花粉育性和裂药指数都较低,变异系数却较大,显示出较丰富的多样性。利用激光扫描共聚焦显微镜术研究发现,高州普通野生稻花粉发育过程与栽培稻一致,同样经历了8个时期,即小孢子母细胞形成期、小袍子母细胞减数分裂期、小孢子早期、小孢子中期、小孢子晚期、二孢花粉期和成熟花粉期。     

The final split: the regulation of anther dehiscence

Controlling male fertility is an important goal for plant reproduction and selective breeding. Hybrid vigour results in superior growth rates and increased yields of hybrids compared with inbred lines; however, hybrid generation is costly and time consuming. A better understanding of anther development and pollen release will provide effective mechanisms for the control of male fertility and for hybrid generation. Male sterility is associated not only with the lack of viable pollen, but also with the failure of pollen release. In such instances a failure of anther dehiscence has the advantage that viable pollen is produced, which can be used for subsequent rescue of fertility. Anther dehiscence is a multistage process involving localized cellular differentiation and degeneration, combined with changes to the structure and water status of the anther to facilitate complete opening and pollen release. After microspore release the anther endothecium undergoes expansion and deposition of ligno-cellulosic secondary thickening. The septum separating the two locules is then enzymatically lysed and undergoes a programmed cell death-like breakdown. The stomium subsequently splits as a consequence of the stresses associated with pollen swelling and anther dehydration. The physical constraints imposed by the thickening in the endothecium limit expansion, placing additional stress on the anther, so as it dehydrates it opens and the pollen is released. Jasmonic acid has been shown to be a critical signal for dehiscence, although other hormones, particularly auxin, are also involved. The key regulators and physical constraints of anther dehiscence are discussed.     

Effect of high temperature and water stress on pollen germination and spikelet fertility in rice

In future climates, rice could more frequently be subjected to simultaneous high temperature and water stress during sensitive developmental stages such as flowering. In this study, five rice genotypes were exposed to high temperature, water stress and combined high temperature and water stress during flowering to quantify their response through spikelet fertility. Microscopic analyses revealed significant differences in anther dehiscence between treatments and genotypes, with a moderately high association with the number of germinated pollen grains on the stigma. There was a strong relationship between spikelet fertility and the number of germinated pollen on stigmas. Although, all three stress treatments resulted in spikelet sterility, high-temperature stress caused the highest sterility in all five genotypes. A cumulative linear decline in spikelet fertility with increasing duration of independent high-temperature stress and in combination with water stress was quantified. Better anther dehiscence, higher in vivo pollen germination, and higher spikelet fertility were observed in both the N22 accessions compared with IR64, Apo and Moroberekan under high temperature, water stress and combined stress, indicating its ability to tolerate multiple abiotic stresses.     

Epidermal jasmonate perception is sufficient for all aspects of jasmonate‐mediated male fertility in Arabidopsis

Jasmonate (JA) signaling is essential for several environmental responses and reproductive development in many plant species. In Arabidopsis thaliana, the most obvious phenotype of JA biosynthetic and perception mutants is profound sporophytic male sterility characterized by failure of stamen filament elongation, severe delay of anther dehiscence and pollen inviability. The site of action of JA in the context of reproductive development has been discussed, but the ideas have not been tested experimentally. To this end we used targeted expression of a COI1‐YFP transgene in the coi1‐1 mutant background. As COI1 is an essential component of the JA co‐receptor complex, the null coi1‐1 mutant is male sterile due to lack of JA perception. We show that expression of COI1‐YFP in the epidermis of the stamen filament and anther in coi1 mutant plants is sufficient to rescue filament elongation, anther dehiscence and pollen viability. In contrast, filament expression alone or expression in the tapetum do not restore dehiscence and pollen viability. These results demonstrate that epidermal JA perception is sufficient for anther function and pollen viability, and suggest the presence of a JA‐dependent non‐autonomous signal produced in the anther epidermis to synchronize both anther dehiscence and pollen maturation.