景宁木兰花粉萌发与贮藏特性研究

以景宁木兰的花粉为试材,采用花粉离体培养法,用单因子与正交试验研究不同浓度蔗糖、H3BO3、CaCl2所组成的基本培养基对景宁木兰花粉萌发的影响,同时探讨不同贮藏条件和贮藏时间对花粉活力的影响。结果表明:景宁木兰在30 g·L-1蔗糖+200 mg·L-1H3BO3+200 mg·L-1CaCl2的液体培养基上萌发率最高(74.56%)。低温条件下有利于景宁木兰花粉生活力的保持,在-80℃条件下,花粉生活力下降较慢,并且随着贮藏时间的增加,经过硅胶干燥的花粉的活力明显高于湿润花粉的活力。     

木兰属(Magnolia)系统发育探讨

在传统的形态解剖基础上 ,结合木兰科 (Magnoliaceae)种子内种皮合点区呈孔状或管状的新的分类依据 ,探讨了木兰属 (Magnolia)的系统发育 ,并提出了木兰属内可能的演化途径。     

香港木兰开花生物学研究

对香港木兰的花果期物候、花朵结构、单花期、开花特性及花粉在离体培养和人工授粉条件下的萌发现象进行观察和研究。结果表明,该种的开花习性限制蜂类昆虫在其柱头可授期内进入花内传粉,为专一的甲虫传粉植物;花粉萌发时间长、开花不集中、花期适逢雨季都影响到昆虫的传粉活动和有效受精。     

景宁木兰组织培养外植体选择与抗褐化研究

该研究以景宁木兰(Magnolia sinostellata)为材料,从外植体类型的选择、消毒时间、预处理方法、离体培养条件和抗褐化剂类型选择等方面进行综合研究,以期筛选出景宁木兰组织培养的最佳方案。结果表明:利用0.1%氯化汞(HgCl_2)浸泡处理14和8 min分别是景宁木兰叶片和带芽茎段、根部的最佳消毒时间;景宁木兰叶片及茎段的褐化率在暗培养7 d时,分别为60.00%和56.67%,14 d时显著升高,而根部则在暗培养14 d时褐化率最低(45%);利用1 g·L~(-1)聚乙烯吡咯烷酮(PVP)浸泡预处理景宁木兰外植体6 h可显著降低(P0.05)其褐化程度,处理后叶片、带芽茎段和根部的褐化率分别降至45.00%、28.33%和63.33%。不同的抗褐化剂均可减轻外植体的褐化程度,但针对不同外植体其效果不同。景宁木兰叶片最佳抗褐化剂为0.02 g·L~(-1)硝酸银(AgNO_3),可使其褐化率降低至16.67%,成活率为11.67%;带芽茎段和根部的最优抗褐化剂为2 g·L~(-1)柠檬酸(CA),褐化率分别降至30.00%和5.00%,成活率依次为50.00%和76.67%。综上所述,带芽茎段和根部抗褐化处理后褐化率较低且成活率较高,为景宁木兰组织培养最佳外植体。     

杂交石竹授粉受精及其胚胎发育过程的观察

以‘杂交石竹’为试验材料,利用荧光显微镜观察其授粉后花粉萌发、花粉管生长情况,采用石蜡切片法对其受精及胚胎发育过程进行观察研究。结果表明:(1)授粉后1h母本柱头上少量花粉开始萌发;授粉后4h大量花粉萌发,花粉管生长至柱头中部有胼胝质出现;授粉后6h花粉管生长至子房组织并有少量与胚珠结合;授粉后15h柱头中出现大量胼胝质,花粉管与胚珠结合数增多;授粉后24h胚珠周围出现多条花粉管,其中1条花粉管进入胚珠,部分进入胚囊的花粉管卷曲盘绕生长并产生胼胝质;精细胞与极核的融合主要发生在授粉后17~48h,与卵细胞融合主要于授粉后1~3d。(2)杂交石竹胚发育经过原胚、球形胚、棒状形胚、心形胚、鱼雷形胚和子叶形胚阶段。(3)杂交障碍表现为:只有游离的胚乳核而无胚发育的胚囊、合子未分裂、两极核未融合、球形胚败育。研究表明,杂交石竹存在受精前和受精后障碍,这是导致其结实率低的主要原因。     

梅与山桃远缘杂交亲和性初步研究

为培育早花抗寒梅花新品种,以梅(Prunus mume)品种‘江梅’(P.mume‘Jiangmei’)、‘淡丰后’(P.mume‘Dan Fenghou’)与山桃(P.davidiana)、‘白花’山桃(P.davidiana‘Alba’)为亲本进行杂交试验,记录种间杂交结实率,观察花粉管生长,对未成熟胚进行培养。结果表明:(1)梅与山桃、‘白花’山桃杂交结实率很低,‘江梅’ב白花’山桃未结果,结实率最高的组合为‘淡丰后’×山桃,也仅有7.4%,且杂交果实的果核内部分胚干瘪、败育。(2)山桃和‘白花’山桃的花粉在梅柱头上都能正常萌发,但花粉管生长受抑制,多数花粉管到达花柱中部即弯曲、缠绕、断裂,花粉管生长过程中有大量的胼胝质产生,表现较低的杂交亲和性,但不同种间杂交亲和程度又有所不同。(3)通过未成熟胚培养获得了杂种苗。研究表明,梅与同属种杂交存在不亲和性,幼胚拯救是获得梅与李属其他种远缘杂交杂种苗的有效途径。     

单性木兰花粉壁超微结构的研究

本文首次报道单性木兰花粉的形态特征和外壁超微结构。花粉外部形态与木兰科各属特征一致,表明单性木兰花粉的原始性。但花粉外壁超微结构显示其外壁表面具皱波状纹饰;覆盖层波浪状,具稀疏的小穿孔;柱状层很薄,小柱典型,直立,短而细。外壁超微结构表明单性木兰属在木兰亚族中是一个较进化的属。扫描电镜和透射电镜资料揭示出单性木兰在植物分类系统中的特殊地位和重要学术价值。     

小报春不同授粉组合亲和性研究

利用小报春‘红星’、‘紫霞’、‘罗兰香’3个品种分别设置8种授粉组合,统计了不同授粉组合的结实情况和单果种子数量,并对‘红星’品种部分授粉组合的花粉萌发与花粉管伸长过程进行荧光显微观察。结果显示:(1)3个品种的结实率和单果种子数量均表现为异型植株授粉>长花柱同型异株授粉、长花柱自交>短花柱同型异株授粉、短花柱自交,不同授粉组合间单果种子数量差异极显著,异型植株授粉类型结实率均达到100%,单果种子数量为44～181粒,显著高于自交组合和同型异株杂交组合;以异型致死花粉作为蒙导对短花柱同型异株授粉组合的结实有一定促进作用,但对长花柱同型异株授粉组合的结实没有一致的促进作用;3个品种中‘红星’的结实率和单果种子数量最高。(2)荧光显微观察表明,异型授粉组合花粉在柱头上大量萌发并在花柱中伸长,授粉144h后花粉管开始进入子房;同型授粉组合授粉12h后花粉开始萌发,但直到授粉后144h花粉管还没有进入花柱;自交授粉组合授粉后144h仍无花粉萌发。实验结果说明,小报春存在明显的自交不亲和性,且短花柱类型的自交不亲和性比长花柱类型强。     

百合远缘杂交花粉萌发及花粉管生长过程观察

利用荧光显微镜对百合远缘杂交组合Bernini×Pollyanna花粉萌发及花粉管生长过程进行观察研究,结果显示,授粉后3~30 h内花粉萌发形成花粉管,且花粉管生长速度由快到慢,48~51 h内花粉管停止生长,花粉管最后深入到花柱的1/3处,并观察到一些异常的花粉管形态;花粉管生长过程中还伴随着一系列的胼胝质反应,出现的部位依次是乳突细胞、花粉管、花柱通道细胞、胚珠中的胚囊.结果表明该杂交组合不亲和.研究认为Berni-ni柱头乳突细胞是阻碍Pollyanna花粉管生长的第一道屏障,花柱通道细胞是抑制花粉管在花柱内生长的第二道屏障.     

烟草脱外壁花粉人工萌发与离体授粉实验系统的建立

通过花蕾低温处理、花药漂浮培养与花粉短时酶解程序可脱去花粉外壁,分离出烟草(Nicotianatabacum L.)的脱外壁花粉。研究了分离过程中的酶液渗透压、培养基中聚乙二醇(PEG)与蔗糖以及添加水解乳蛋白等因素对脱外壁花粉人工萌发的影响。在含30％PEG-6000与0.1％水解乳蛋白的D_2培养基中,萌发率最高达57.8％;花粉管生长正常,培养24h后一半以上的花粉管中生殖细胞分裂成精子。用微滴和贴滤纸小片的方法将30～40粒脱外壁花粉授予柱头上,近一半能萌发花粉管并在花柱中生长。采取授粉后子房培养方法,获得了种子与幼苗。从而建立了脱外壁花粉离体授粉实验系统。讨论了脱外壁花粉人工萌发与离体授粉实验系统的建立对于研究外壁在花粉萌发中的生物学功能以及开拓新的转基因技术等方面的意义。     

梨远缘花粉原位萌发及生长特性

应用荧光标记方法对梨远缘花粉在‘丰水’和‘噢嗄二十世纪’柱头上萌发及花粉管生长特性进行观察,结果表明:(1)梨远缘花粉均能在柱头上萌发,但其萌发率不同,授粉后24 h,在‘丰水’柱头上‘红叶桃’花粉萌发率最高,达62.8%,而‘盖县大李’花粉萌发率仅为12.0%,各种远缘花粉在‘丰水’柱头萌发率均高于‘噢嗄二十世纪’柱头。(2)各种远缘花粉管在梨柱头或花柱内生长情况也有差异,‘红叶桃’等核果类花粉管在梨柱头上均表现为扭曲、盘绕等现象,不能穿过柱头;‘红星’和‘红富士’花粉管虽然有少量穿过柱头,但不能进一步在花柱内生长,表现为扭曲变形、先端膨大等不亲和性现象。因此,梨与远缘果树杂交不亲和在柱头上就已发生,这与梨自交不亲和反应发生在花柱内的现象不同。     

山玉兰花粉离体萌发和花粉管生长特性的研究

山玉兰(Magnolia delavayi)是木兰科木兰属的常绿乔木或大型灌木,是重要的园林造景、庭院绿化素材,也是重要的育种资源。山玉兰花粉的研究对其杂交育种的成败具有重要影响,但目前尚未见其花粉活力的相关报道。该研究以新鲜的山玉兰花粉为对象,采用悬滴培养法分析了温度、光照以及培养液的蔗糖和硼酸浓度对山玉兰花粉萌发的影响。结果表明:(1)山玉兰花粉萌发时,最适宜的温度为27℃。(2)光暗条件下,山玉兰花粉以浓度为5%的蔗糖培养效果最佳,其萌发率在16%以上;而硼酸浓度则以0.001%的培养效果最佳。(3)蔗糖与硼酸共同作用可有效促进花粉萌发和花粉管生长。其中,在光照条件下,以5.0%蔗糖+0.001%硼酸为最适宜的培养液,花粉萌发率达41.27%,花粉管长达281.49μm;而在黑暗条件下,则以5.0%蔗糖+0.01%硼酸为最适宜的培养液,花粉萌发率达45.71%,花粉管长达254.00μm。该研究结果为进一步开展人工辅助授粉、发掘山玉兰的种质资源工作奠定了基础。     

Observation of Flowering Process of Grape (<i>Vitis vinifera</i> L.)—Insight into the Starting of Pollination of Flower Hiding in Calyptras (Cap)

It is generally agreed that many Vitis vinifera L. cultivars are self-fertile, where self-pollination often occurs before capfall in a process called cleistogamy. Therefore, it is difficult to identify the right time to remove stamens before self-pollination during the cross-breeding of grape. For this paper, we observed the process of grape flowering and measured the pollen viability and stigma receptivity of grape flowers of ‘Shine Muscat’ in order to identify the starting time of self-pollination before capfall and to provide useful information for improving the efficiency of cross-breeding. The results demonstrate that the anther is not cracked during the visible clusters and separated clusters stages. Meanwhile, in the separated floral buds, flowering begins, and full bloom stages, the pollen viability is 60.7%, 73.2% and 80.3%, respectively; however, at the berry set stage, pollen viability drops to zero. The top of the mature stigma is composed of a layer of nearly cylindrical papillary cells, and the stigma receptivity for pollen changes with the development of flowers: in particular, no reaction was observed in the visible clusters stage; weak positive reaction at the separated clusters stage; strong positive reaction at the separated floral buds, flowering begins, and full bloom stages; and no reaction at the berry set stage. In the separated floral buds stage, pollen tubes were seen germinating in the style. In the flowering begins stage, more pollen tubes were observed at the entry of the ovary. During the full bloom stage, most pollen tubes elongated into the ovary base and some entered the pearl hole. At the berry set stage, newborn endosperm nucleus could be seen in the ovule. From the above, we can conclude that the initiation time of closed fertilization for ‘Shine Muscat’ grape can be judged as the separated floral buds stage, and it is best to discard the stamen before the separated floral buds stage when conducting cross-breeding.     

Effects of pollen shortage and self-pollination on seed production of an endangered tree, Magnolia stellata

BACKGROUND AND AIMS: Pollen limitation is a significant determinant of seed production, and can result from both insufficient pollen quantity (pollen shortage) and quality (mainly relating to self-pollination). For animal-pollinated tree species with large floral displays, pollen limitation may be determined by a balance between increased pollen quantity due to increased attractiveness for pollinators, countered by increased self-pollination due to increased geitonogamy. The contributions of pollen shortage and self-pollination on seed production were quantitatively examined in the natural pollination of an insect-pollinated, dichogamous, endangered tree, Magnolia stellata, which has a large, showy floral display. METHODS: Manual self- and cross-pollinations were conducted to determine the effects of selfing on seed production. The outcrossing rate was measured using microsatellite analyses of open-pollinated seeds, and the embryo mortality rate caused by self-pollination was indirectly estimated. The frequency of ovule mortality due to pollen shortage was also inferred using the embryo mortality and ovule survival rates from natural pollination. KEY RESULTS: The average fruit set, seed set per fruit, and ovule survival rate per tree from hand cross-pollination were 1.37, 3.15, and 3.34 times higher than those from hand self-pollination, respectively, indicating that self-pollination causes inbreeding depression for fruit and seed set. The multilocus-outcrossing rate (t(m)) was intermediate, 0.632, and the primary selfing rate was 0.657. This indicates that frequent geitonogamous selfing occurs. The ovule mortality rate due to pollen shortage and the embryo mortality rate due to self-pollination were estimated to be 80.8 % and 45.9 %, respectively. CONCLUSIONS: It is concluded that seed production of M. stellata is strongly limited by both pollen shortage and self-pollination. Inefficient beetle-pollination and the automimicry system via asynchronous flowering might be responsible for the high level of pollen shortage and frequent geitonogamy. This is despite a large, showy floral display and the dichogamous system of the species.     

Reproductive and Pollination Biology of Magnolia and its Allied Genera (Magnoliaceae). I. Floral Volatiles of Several Magnolia and Michelia Species and their Roles in Attracting Insects

Abstract Volatile substances emitted from the flowers of eight Magnolia taxa ( M. sieboldii ssp. japonica, M. praecocissima var. praecocissima and var. borealis, M. tomentosa, M. salicifolia, M. obovata, M. denudata, and M. grandiflora ) and one Michelia species ( M. compressa ) (Magnoliaceae) were examined and identified using GC-MS. Volatile substances of these Magnolia and Michelia species consist primarily of monoterpenoids and sesquiterpenoids produced by the mevalonate pathway, acetogenins by the acetate-malonate pathway, and phenyl-propanoids by the shikimate pathway. These Magnolia and Michelia species all possessed various combinations of volatile monoterpenoids, acetogenins, and phenylpropanoids, except for Magnolia obovata , which emitted primarily sesquiterpenoids. Free amino acids in pollen of 12 Magnolia and one Liriodendron species were also analyzed, and their value as food sources for pollinators evaluated.
Pollinators visiting the flowers of five Magnolia species were collected in their native sites and identified. Their behaviors and roles as pollinating agents were assessed.     

红豆杉的胚珠发育,传粉滴形成和传粉过程

观察了红豆杉（Ｔａｘｕｓ ｃｈｉｎｅｎｓｉｓ （Ｐｉｌｇ．）Ｒｅｈｄ）的花粉形态和水合特性及胚珠发育、传粉滴形成与传粉过程。成熟花粉为单细胞,无气吓,形状不规则,外壁表面具大量乌氏体。花粉水合时,内壁膨胀,外壁开裂。通常情况下,外壁保留在水滴或传粉滴的表面,而花粉的其他部分进入水滴或传粉滴内。在８月下旬,可观察到下弯的雌性生殖芽。下弯这一特性是雌性生殖芽区别于营养芽的重要特征。这一时期的雌性生殖芽