Pollen Grain Germination and Pollen Tube Growth in Pistil of Rice

The germination of pollen grain in vitro and the growth of pollen tube in the pistil of rice were observed with a microscope. The stigma was removed at different time points after pollination to study its effect on seed setting rate. The rice pollen grain started to germinate at 2 min after pollination and the pollen tube penetrated stigma into style in 5-10 min, 30 min later the end of pollen tube reached the bottom of ovary, and only some pollen tubes arrived at embryo sac at 40 min after pollination. Meanwhile, a small amount of callose began to deposit in the pollen tubes, a great deal of callose was observed at 50 min after pollination, whereas the pollen grain began to shrink. The growing rates of pollen tube in the rice stigma, style and ovary were 1500, 5000, and 5400 μm/h, respectively. The seed setting rate was quite low when the stigma was removed at about 10-15 min after pollination, gradually increased when it removed at 20 min to 50 min after pollination, and over 60% when it removed at 50 min after pollination and finally tended to be stable.

Rice pollen-specific OsRALF17 and OsRALF19 are essential for pollen tube growth

Pollen tube growth is essential for successful double fertilization, which is critical for grain yield in crop plants. Rapid alkalinization factors(RALFs) function as ligands for signal transduction during fertilization. However, functional studies on RALF in monocot plants are lacking.Herein, we functionally characterized two pollen-specific RALFs in rice(Oryza sativa) using multiple clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9-induced loss-of-function mutants,peptide treatment, expression analyses, and tag reporter lines. Among the 41 RALF members in rice, OsRALF17 was specifically expressed at the highest level in pollen and pollen tubes.Exogenously applied OsRALF17 or OsRALF19peptide inhibited pollen tube germination and elongation at high concentrations but enhanced tube elongation at low concentrations, indicating growth regulation. Double mutants of OsRALF17 and OsRALF19(ralf17/19) exhibited almost full male sterility with defects in pollen hydration, germination, and tube elongation,which was partially recovered by exogenous treatment with OsRALF17 peptide. This study revealed that two partially functionally redundant OsRALF17 and OsRALF19 bind to Oryza sativa male-gene transfer defective 2(OsMTD2) and transmit reactive oxygen species signals for pollen tube germination and integrity maintenance in rice. Transcriptomic analysis confirmed their common downstream genes, in osmtd2 and ralf17/19. This study provides new insights into the role of RALF, expanding our knowledge of the biological role of RALF in regulating rice fertilization.

Screening for High-Temperature Tolerant Cotton Cultivars by Testing In Vitro Pollen Germination,Pollen Tube Growth and Boll Retention

With radical global climate change and global warming, high temperature stress has become one of major factors exerting a major Influence on crop production, In the cotton （Gossyplum hirsutum L.）-growlng areas of China, especially in the Yangtze River valley, unexpected periodic episodes of extreme heat stress usually occur In July and August, the peak time of cotton flowering and boll loading, resulting In lower boll set and lint yield. Breeding programs for screening high temperature-tolerant cotton germplasm and cultlvars are urgent In order to stabilize yield in the current and future warmer weather conditions. In the present study, 14 cotton cultivars were quantified for in vitro pollen germination and pollen tube growth in response to temperatures ranging from 10 to 50 ℃ at 5 ℃ intervals. Different cotton genotypes varied In their in vitro pollen germination and pollen tube length responses to the different temperatures. Maximum pollen germination and pollen tube length ranged from 25.2% to 56.2% and from 414 to 682 pro, respectively. The average cardinal temperatures （Tmin,, Topt, and Tmax） also varied among the 14 cultivars and were 11.8, 27.3, and 42.7 ℃ for pollen germination and 11.8, 27.8, and 44.1 ℃ for maximum pollen tube length. Variations In boll retention and boll numbers per plant in field experiments were found for the 14 cotton cultivars and the boll retention and boll retained per plant on 20 August varied considerably In different years according to weather conditions. Boll retention on 20 August was highly correlated with maximum pollen germination （R^2 = 0.84） and pollen tube length （R^2=0.64）. A screening method based on principle component analysis of the combination of pollen characterlatics In an in vitro experiment and boll retention testing In the field environment was used In the present study and, as a result, the 14 cotton cultlvars could be classified as tolerant, moderately tolerant, moderately susceptible and susceptible to high temperature.

The Calcineurin B-Like Ca2＋ Sensors CBL1 and CBL9 Function in Pollen Germination and Pollen Tube Growth in Arabidopsis

Ca2＋ has been established as an important second messenger regulating pollen germination and tube growth. However, to date, only a few signaling components have been identified to decode and relay Ca2＋ signals in growing pollen tubes. Here, we report a function for the calcineurin B-like （CBL） Ca2＋ sensor proteins CBL1 and CBL9 from Arabidopsis in pollen germination and tube growth. Both proteins are expressed in mature pollen and pollen tubes and impair pollen tube growth and morphology if transiently overexpressed in tobacco pollen. The induction of these phenotypes requires efficient plasma membrane targeting of CBL1 and is independent of Ca2＋ binding to the fourth EF-hand of CBL1. Overexpression of CBL1 or its closest homolog CBL9 in Arabidopsis renders pollen germination and tube growth hypersensitive towards high external K＋ concentrations while disruption of CBL1 and CBL9 reduces pollen tube growth under low K~ conditions. Together, our data identify a crucial function for CBL1 and CBL9 in pollen germination and tube growth and suggest a model in which both proteins act at the plasma membrane through regulation of K＋ homeostasis.

Higher-Ordered Actin Structures Remodeled by Arabidopsis ACTIN-DEPOLYMERIZING FACTOR5 Are Important for Pollen Germination and Pollen Tube Growth

Dynamics of the actin cytoskeleton are essential for pollen germination and pollen tube growth. ACTIN- DEPOLYMERIZlNG FACTORs （ADFs） typically contribute to actin turnover by severing/depolymerizing actin filaments. Recently, we demonstrated that Arabidopsis subclass III ADFs （ADF5 and ADF9） evolved F-actin-bundling function from conserved F-actin-depolymerizing function. However, little is known about the physiological function, the evolutional significance, and the actin-bundling mechanism of these neo- functionalized ADFs. Here, we report that loss of ADF5 function caused delayed pollen germination, retarded pollen tube growth, and increased sensitive to latrunculin B （LatB） treatment by affecting the generation and maintenance of actin bundles. Examination of actin filament dynamics in living cells revealed that the bundling frequency was significantly decreased in adf5 pollen tubes, consistent with its biochem- ical functions. Further biochemical and genetic complementation analyses demonstrated that both the N- and C-terminal actin-binding domains of ADF5 are required for its physiological and biochemical functions. Interestingly, while both are atypical actin-bundling ADFs, ADF5, but not ADF9, plays an important role in mature pollen physiological activities. Taken together, our results suggest that ADF5 has evolved the function of bundling actin filaments and plays an important role in the formation, organization, and maintenance of actin bundles during pollen germination and pollen tube growth.

Expression of an Exo-β-glucanase Gene Is Specifically Related to Lily Pollen Germination and Pollen Tube Growth

The present study is expected to provide better understanding of how exo-β-glucanase acts on the biochemo-rheological property of the cell wall, and thus the regulation mechanism of pollen tube growth. A polymerase chain reaction （PCR）-based strategy and cDNA library screening were used to clone an exo-β-glucanase gene （LP-Exol） from Lilium Iongiflorum pollen tubes. The LP-Exol sequence showed a high level of homology to that of reported plant exo-β-glucan hydrolases. Phylogenetic analysis demonstrated that there was close relationship between LP-Exol and other exo-glucan hydrolases in Poacea plants. Northern analysis indicated that LP-Exol transcripts stored in pollen grains at low levels; LP-Exol transcription level enhanced at 2 h after the onset of pollen incubation and remained relatively high during pollen tube elongation. Moreover, the expression of LP-Exol was undetectable in petals, filaments, stigmas, styles, ovaries, leaves, and stems. These results suggest that LP-Exol gene is a late pollen gene, specifically contributing to the regulation of lily pollen germination and pollen tube growth.

Pollen Tube Growth: a Delicate Equilibrium Between Secretory and Endocytic Pathways

Although pollen tube growth is a prerequisite for higher plant fertilization and seed production, the processes leading to pollen tube emission and elongation are crucial for understanding the basic mechanisms of tip growth. It was generally accepted that pollen tube elongation occurs by accumulation and fusion of Golgi-derived secretory vesicles （SVs） in the apical region, or clear zone, where they were thought to fuse with a restricted area of the apical plasma membrane （PM）, defining the apical growth domain. Fusion of SVs at the tip reverses outside cell wall material and provides new segments of PM. However, electron microscopy studies have clearly shown that the PM incorporated at the tip greatly exceeds elongation and a mechanism of PM retrieval was already postulated in the mid-nineteenth century. Recent studies on endocytosis during pollen tube growth showed that different endocytic pathways occurred in distinct zones of the tube, including the apex, and led to a new hypothesis to explain vesicle accumulation at the tip; namely, that endocytic vesicles contribute substantially to V-shaped vesicle accumulation in addition to SVs and that exocytosis does not involve the entire apical domain. New insights suggested the intriguing hypothesis that modulation between exo- and endocytosis in the apex contributes to maintain PM polarity in terms of lipid/protein composition and showed distinct degradation pathways that could have different functions in the physiology of the cell. Pollen tube growth in vivo is closely regulated by interaction with style molecules. The study of endocytosis and membrane recycling in pollen tubes opens new perspectives to studying pollen tube-style interactions in vivo.

Characterization of an endo-1,4-β-glucanase from Lilium longiflorum that functioned in pollen germination and pollen tube growth

Endo-β-glucanases play vital roles in the regulation of pollen tube growth. Here, a previously identified endo-1,4-β-glucanase from Lilium longiflorum (lily), named LlpCel1, was expressed in Escherichia coli, purified, and further investigated for its physiological function. The recombinant LlpCel1 protein hydrolyzed carboxy-methylcellulose (CMC) and exhibited activity towards laminarin from Eisenia. arborea and 1,3:1,4-β-glucan of barley. The pH for the optimum activity was 6.0 and the value of Km calculated from CMC was 5.0 mg/mL. Adding EDTA resulted in the total loss of the enzymatic activity, and this effect could be restored by the addition of Ca2+. Western blotting analysis showed that LlpCel1 protein was present in pollen grains and rehydrated pollen grains, and the amount of the protein was increased during pollen germinating, but not in the pollen tube. Consistently, the immunofluorescence labeling study with the antibody against LlpCel1 also indicated the presence of LlpCel1 at the begin-ning of germination, but not in the elongating pollen tube. Furthermore, incubation of LlpCel1 with pollen at the beginning of pollen germination increased the germination percentage and the length of pollen tube. All of these results suggested that LlpCel1 could play an important role in the regulation of lily pollen germination and the initiation of pollen tube growth.

AGC1.5 Kinase Phosphorylates RopGEFs to Control Pollen Tube Growth

Double fertilization in angiosperms requires the targeted delivery of immotile sperm to the eggs through pollen tubes. The polarity of tip-growing pollen tubes is maintained through dynamic association of active Rho GTPases of plants （ROP-GTP） with the apical plasma membrane. Guanine nucleotide exchange factors for ROPs （RopGEFs） catalyze the activation of ROPs and thereby affect spatiotemporal ROP signaling. Whereas RopGEFs have been found to be phosphorylated proteins, the kinases responsible for their phos- phorylation in v/vo and biological consequences of RopGEF phosphorylation in pollen tube growth remain unclear. We report here that the Arabidopsis AGC1.5 subfamily of cytoplasmic kinases is critical for the restricted localization of ROP-GTP during pollen tube growth. Loss of AGCI.5 and AGC1.7 functions resulted in the mistargeting of active ROPs and defective events downstream of ROP signaling in pollen tubes. AGCI.5 interacts with RopGEFs via their catalytic PRONE domain and phosphorylates RopGEFs at a conserved Ser residue of PRONE domain. Loss of AGC1.5 and AGC1.7 functions resulted in the mistargeting of RopGEFs in pollen tubes, similar to the phenotype caused by the mutation that renders RopGEFs non-phosphorylatable by AGC1.5. Collectively, our results provide mechanistic insights into the spatiotem- poral activation of ROPs during the polar growth of pollen tubes.

Inositol (1,4,5)-triphosphate Modulates Pollen Tube Polar Growth

In this article, the functions of D-myo-inositol-1,4,5-trisphosphate (Ins [1,4,5] P3) in regulating pollen tube polar growth were investigated by application of caged version of the phosphoinositides. To increase the intracellular Ins（1,4,5）P3 concentration at the apical region of pollen tube, the caged Ins（1,4,5）P3 loaded by osmotic shock was activated by 10 s 360 nm UV flash at this domain （10 μm from the tip）. More than 70% pollen tubes were induced swelling at apex and/or growth axis reorientation, accompanying temporarily growth arrest, by localized increase of Ins（1,4,5）P3 concentration （n=21）. While, pollen tubes without being loaded with caged Ins（1,4,5）P3 had not response to the same dosage UV flash. With FM 1-43 fluorescent staining, it was found that growth perturbation by UV activated caged Ins（1,4,5）P3 had tight link with membrane trafficking at the apical zone of pollen tubes. Upon UV pulse, the apical V-shaped bright area where was full of secretory vesicles spread to a much broader region, which implied that actin filaments at the apical region were remodeled. It was also observed that the FM 1-43 fluorescence intensity at tip remarkably increased than that before UV flash, which demonstrated that more secretory vesicles were accumulated at this region. To estimate the role of Ins（1,4,5）P3 in modulating intracellular calcium concentration and distribution, dextran conjugated fluorescent dye Calcium Green-1 and Rhodamine B were microinjected into pollen tubes together with caged Ins（1,4,5）P3. The results showed that calcium concentration at the subapical region increased upon UV released Ins（1,4,5）P3. Consequently, the tip-focused calcium gradient under the apical dome of pollen tube was disturbed. Simultaneously, the pollen tube bulged at the apical region and its growth rate decreased. As the tip-focused calcium gradient was reestablished, the pollen tube morphology and growth recovered to the normal level. Therefore, Ins（1,4,5）P3 can modulate pollen tube growth rate and direction through mobilizing intracellular calcium and regulating vesicle trafficking during pollen tube finding its way to the ovary.

Self S‑RNase reduces the expression of two pollen‑specific COBRA genes to inhibit pollen tube growth in pear

Due to self-incompatibility(SI)prevents self-fertilization,natural or artificial cross-pollination has been conducted in many orchards to stabilize fruit yield.However,it is still puzzled which routes of self S-RNase arresting pollen tube growth.Herein,17 COBRA genes were isolated from pear genome.Of these genes,the pollen-specifically expressed PbCOB.A.1 and PbCOB.A.2 positively mediates pollen tube growth.The promoters of PbCOB.A.1 and/or PbCOB.A.2 were bound and activated by PbABF.E.2(an ABRE-binding factor)and PbC2H2.K16.2(a C2H2-type zinc finger protein).Notably,the expressions of PbCOB.A.1,PbCOB.A.2,and PbC2H2.K16.2 were repressed by self S-RNase,suggesting that self S-RNase reduces the expression of PbCOB.A.1 and PbCOB.A.2 by decreasing the expression of their upstream factors,such as PbC2H2.K16.2,to arrest pollen tube growth.PbCOB.A.1 or PbCOB.A.2 accelerates the growth of pollen tubes treated by self S-RNase,but can hardly affect level of reactive oxygen species and deploymerization of actin cytoskeleton in pollen tubes and cannot physically interact with any reported proteins involved in SI.These results indicate that PbCOB.A.1 and PbCOB.A.2 may not relieve S-RNase toxicity in incompatible pollen tube.The information provides a new route to elucidate the arresting pollen tube growth during SI reaction.

攸县油茶远缘杂交亲和性的解剖学研究

【目的】为了探究攸县油茶(Camellia yuhsienensis)远缘杂交亲和性的生物学机制。【方法】研究以石果毛蕊山茶[C.mairei var.lapidea(Y.C.Wu)Sealy](SG)、海南油茶‘海油3号’(C.hainanica‘Haiyou 3’)(HN-3)和普通油茶‘德油6号’(C.oleifera‘Deyou 6’)(DY-6)为父本,采用常规授粉和混合授粉方式对攸县油茶(YX)进行自交和杂交授粉。采用流式细胞仪测定亲本倍性,通过离体萌发法测定父本花粉活力,运用显微技术观察花粉萌发及花粉管生长动态以及早期果实和胚珠发育动态,并田间调查各授粉组合的座果率和落果率。【结果】(1)YX为六倍体,SG为二倍体,HN-3为八倍体,DY-6为六倍体。(2)4种油茶花粉具有较强的活力和育性,其中花粉活力最高的是DY-6,为70.13%;花粉活力最低的是YX,为40.68%。(3)自交和杂交所有组合的花粉均能在柱头上正常萌发,且花粉管在授粉后12 h均生长到花柱基部,但自交和YX×SG的花粉管进入子房后生长受阻,无法到达胚珠;YX×HN-3和YX×DY-6的花粉管则在授粉后72h到达胚珠。(4)自交和杂交授粉后100 d内,YX×DY-6的果实横径、纵径最大,YX×SG的果实横径、纵径最小;YX×DY-6的败育胚珠数最少,平均为3.83枚/果;YX×SG的败育胚珠数最多,平均为6.00枚/果。(5)自交和杂交授粉后100 d内,YX×DY-6的座果率最高,为68.0%;YX×SG的座果率最低,为4.0%;YX×HN-3的座果率为16.0%,其落果高峰主要集中在授粉后0~30 d和40~70 d;混合授粉组合YX×SD的座果率为22.0%。【结论】自交和YX×SG的杂交障碍主要为受精前障碍,YX×HN-3的杂交障碍为受精前和受精后障碍;攸县油茶远缘杂交亲和性也与亲本倍性有关,其亲和性大小为YX(六倍体)×DY-6(六倍体)>YX×HN-3(八倍体)>YX×YX>YX×SG(二倍体);混合授粉方式能在一定程度上提高攸县油茶远缘杂交亲和性。研究结果为攸县油茶远缘杂交亲和性机理研究提供了细胞学基础,也为油茶远缘杂交育种工作提供理论参考。

Influence of Storage Time on Pollen Traits in Actinidia eriantha

We examined the influence of storage time on germinability and tube growth of freeze stored pollens collected from 25 wild male plants in Actinidia eriantha variety. Pollens were stored in freezer at - 20°C for six months and one year periods to determine changing at germinability in time. In vitro germination was conducted in certain cultural medium defined for Actinidia genus. The results showed that the germination percentages and tube lengths of genotypes decreased at the end of storage period. MH22, MH45, MH47, MH56, MH67, MH70, MH71, MH72, MH74, MH55 and MH61 genotypes were evaluated as vigor genotypes, because they maintained their viability and germination capability displaying statistically insignificant decreasing although their tube lengths significantly decreased except MH67. This investigation provided to determine some robust wild male germplasm resources in A. eriantha in point of durability of pollens against long term conservation for using at future pollination and breeding programs.

Arabidopsis pollen-specific glycerophosphodiester phosphodiesterase-like genes are essential for pollen tube tip growth

In angiosperms,pollen tube growth is critical for double fertilization and seed formation.Many of the factors involved in pollen tube tip growth are unknown.Here,we report the roles of pollenspecific GLYCEROPHOSPHODIESTER PHOSPHO DIESTERASE-LIKE(GDPD-LIKE)genes in pollen tube tip growth.Arabidopsis thaliana GDPD-LIKE6(At GDPDL6)and At GDPDL7 were specifically expressed in mature pollen grains and pollen tubes and green fluorescent protein(GFP)-At GDPDL6 and GFP-At GDPDL7 fusion proteins were enriched at the plasma membrane at the apex of forming pollen tubes.Atgdpdl6 Atgdpdl7 double mutants displayed severe sterility that was rescued by genetic complementation with At GDPDL6 or At GDPDL7.This sterility was associated with defective male gametophytic transmission.Atgdpdl6 Atgdpdl7 pollen tubes burst immediately after initiation of pollen germination in vitro and in vivo,consistent with the thin and fragile walls in their tips.Cellulose deposition was greatly reduced along the mutant pollen tube tip walls,and the localization of pollen-specific CELLULOSE SYNTHASE-LIKE D1(CSLD1)and CSLD4 was impaired to the apex of mutant pollen tubes.A rice pollen-specific GDPD-LIKE protein also contributed to pollen tube tip growth,suggesting that members of this family have conserved functions in angiosperms.Thus,pollen-specific GDPDLIKEs mediate pollen tube tip growth,possibly by modulating cellulose deposition in pollen tube walls.

自花授粉花柱提取物对澳洲坚果品种间杂交亲和性的影响

【目的】明确澳洲坚果品种间杂交的亲和性,为快速鉴定品种间杂交亲和程度和授粉品种配置提供理论依据和实践支撑。【方法】以澳洲坚果13个品种为母本试材,10个品种为父本试材,设置115个杂交组合,利用母本自交花柱提取液培养基培养父本花粉,测定杂交组合的花粉萌发率及花粉管生长情况,结合花粉萌发受影响程度指数(pollen germination,PG)与花粉管生长受影响程度指数(pollen tube growth,PTG),判定品种间杂交亲和性程度。【结果】明确了115个杂交组合的亲和性程度,筛选出54个强杂交亲和性组合,其中有9对杂交组合可互为亲本;另外筛选出28个中等杂交亲和性组合和33个弱杂交亲和性组合。品种HAES863、O.C、HAES820可以作为杂交育种父本材料,在果园构建中可以作为13个参试母本的授粉树;品种HAES863、D4、HAES951、HAES816、HAES842为极好的杂交育种母本材料。【结论】建立了澳洲坚果品种间杂交亲和性室内鉴定标准,将杂交亲和性划分为强(PG值≤0.5,PTG值≤0.2)、中(0.5<PG值≤0.7;0.2<PTG值≤0.4)和弱(0.7<PG值<1;0.4<PTG值<1)3种类型,用于澳洲坚果杂交亲和性的快速鉴定,为澳洲坚果种植园品种配置提供参考。

爆杖花和红马银花的杂交亲和性分析

以我国特有的爆杖花和易危的红马银花2种杜鹃物种为亲本进行杂交试验,探讨了不同杜鹃亚属间的杂交亲和性,以期为合理利用我国特有和濒危的杜鹃花种质提供依据。测定了2种杜鹃的新鲜花粉活力,并开展2种杜鹃的自交及正反交授粉试验,同时利用荧光显微镜对各杂交组合授粉后0、12、24和96 h的杂交花粉萌发和花粉管生长进行观测,比较了其杂交亲和性。结果显示:爆杖花和红马银花的新鲜花粉活力分别为40.61%和31.43%,均适宜授粉;在红马银花×红马银花、红马银花×爆杖花、爆杖花×爆杖花、爆杖花×红马银花等4个杂交组合中,杂交花粉在授粉后0和12 h时均未萌发,而在授粉后24 h开始萌发;在授粉后96 h,各杂交组合的花粉萌发率和花粉管长度均达到最高值,其中以红马银花×红马银花的花粉萌发率最高,达24.95%,而以爆杖花×红马银花的花粉管最长,为0.93 cm,达到花柱中部。由此可知,爆杖花和红马银花自交亲和,2种杜鹃杂交具有部分亲和性,且爆杖花×红马银花的亲和性好于红马银花×爆杖花。

植物激素对烟草花粉萌发及授粉后花粉管生长的影响

采用离体萌发法研究外源植物激素(GA、IAA、ABA、6-BA)对栽培烟草、花烟草和黄花烟草花粉萌发的影响。结果表明,ABA抑制3个烟草种的花粉萌发;栽培烟草花粉萌发最适的GA浓度为10 mmol/L,IAA浓度为10 mmol/L,6-BA浓度为15 mmol/L。黄花烟草花粉萌发的最适GA浓度为100 mmol/L、IAA浓度为30 mmol/L、6-BA浓度为4 mmol/L。花烟草花粉萌发最适6-BA浓度为1 mmol/L,GA和IAA抑制其花粉萌发。用半离体培养实验探究上述激素对栽培烟草自交,及其作为母本与花烟草和黄花烟草杂交授粉后花粉管生长的影响,结果表明,4种激素均促进花粉管生长,且对栽培烟草(自交亲和)花粉管生长的促进作用最强,但GA抑制花烟草花粉管生长。此外,6-BA还能改善黄花烟草花粉管在杂交授粉过程中的卷曲现象。

阿拉伯半乳糖蛋白在茉莉花粉萌发和花粉管生长中的作用研究

阿拉伯半乳糖蛋白(arabinogalactan proteins,AGPs),是一类具有繁杂构造的糖蛋白,遍及于植物的不同组织中.大量研究表明,AGPs在花粉的黏附、水合、萌发和花粉管在花柱内极性生长过程中都起着重要作用.采用花粉离体培养技术,对AGPs与其抑制剂β葡萄糖基-Yariv试剂(β-D-glucosyl-Yariv reagent,βGlcY)结合后,茉莉花粉萌发和花粉管生长的情况,以及去除抑制剂后,花粉管再生情况进行了研究,以探明AGPs是否对茉莉花粉萌发和花粉管生长起作用.结果表明:Yariv试剂在处理后1~2h内能明显抑制茉莉花粉萌发和花粉管的生长,花粉萌发率和花粉管生长速度与对照相比都显著下降;去除抑制剂后,茉莉花粉管可恢复正常生长能力,故AGPs在茉莉花粉萌发和花粉管生长过程中起着重要的促进作用.研究结果对提高茉莉花粉活力、促进花粉管生长以克服结实障碍、提高结实率有重要的指导意义.

铁甲秋海棠花粉离体萌发研究

为探究铁甲秋海棠Begonia masoniana花粉离体萌发适宜条件,采用液体培养法,通过单因素试验考察蔗糖、硼酸、PEG-4000、氯化钙浓度以及不同培养时间对花粉管生长的影响。结果表明,铁甲秋海棠花粉离体萌发的最适培养基为5%蔗糖+0.01%硼酸+5%PEG-4000+0.003%氯化钙;最佳观察时间为120 min。利用该培养基进行培养,铁甲秋海棠花粉萌发率达90.86%,花粉管长度达511.51μm。该研究结果可为同属其他种类秋海棠的花粉研究及育种提供参考。

Arabidopsis PRK6 interacts specifically with AtRopGEF8/12 and induces depolarized growth of pollen tubes when overexpressed

The pollen receptor kinases (PRK) are critical regulators of pollen tube growth. The Arabidopsis genome encodes eight PRK genes, of which six are highly expressed in pollen tubes. The potential functions of AtPRK1 through AtPRK5, but not of AtPRK6,in pollen growth were analyzed in tobacco. Herein, AtPRK6 was cloned, and its function was identified. AtPRK6 was expressed specifically in pollen tubes. A yeast two-hybrid screen of AtPRK6 against 14 Arabidopsis Rop guanine nucleotide exchange factors (RopGEFs) showed that AtPRK6 interacted with AtRopGEF8 and AtRopGEF12. These interactions were confirmed in Arabidopsis mesophyll protoplasts. The interactions between AtPRK6 and AtRopGEF8/12 were mediated by the C-termini of AtRopGEF8/12 and by the juxtamembrane and kinase domain of AtPRK6, but were not dependent on the kinase activity. In addition, transient overexpression of AtPRK6::GFP in Arabidopsis protoplasts revealed that AtPRK6 was localized to the plasma membrane. Tobacco pollen tubes overexpressing AtPRK6 exhibited shorter tubes with enlarged tips. This depolarized tube growth required the kinase domain of AtPRK6 and was not dependent on kinase activity. Taken together, the results show that AtPRK6,through its juxtamembrane and kinase domains (KD), interacts with AtRopGEF8/12 and plays crucial roles in polarized growth of pollen tubes.