蜜柚不同砧穗组合苗期嫁接亲和性评价

为评价蜜柚砧穗的嫁接亲和性,以红绵蜜柚(Citrus grandis‘Hongmianmiyou’)、三红蜜柚(‘Sanhongmiyou’)、红肉蜜柚(‘Hongroumiyou’)、黄金蜜柚(‘Huangjinmiyou’)和琯溪蜜柚(‘Guanximiyou’)作接穗,枳(Poncirus trifoliata)、香橙(Citrus junos)、酸柚(Citrus grandis)作砧木,田间调查15个砧穗组合苗期生长指标,测定嫁接愈合期叶片多酚氧化酶(PPO)和过氧化物酶(POD)活性、可溶性蛋白和可溶性糖含量,采用主成分分析和聚类分析方法对蜜柚砧穗组合嫁接亲和性进行评价。结果表明,以柚作砧木的砧穗组合保存率高、生长势旺盛、抽梢能力强,以枳和香橙作砧木的砧穗组合部分指标存在差异,其中红绵蜜柚和黄金蜜柚以枳作砧木时表现出不亲和现象。不同砧穗组合嫁接愈合时期PPO、POD、可溶性蛋白和可溶性糖变化趋势基本一致。主成分分析结果表明,4个主成分基本反映了15个指标91.33%的数据信息。聚类分析将15个砧穗组合分为4类,与主成分分析结果基本一致。因此,琯溪蜜柚、红肉蜜柚和三红蜜柚嫁接可采用枳和柚作砧木,红绵蜜柚和黄金蜜柚嫁接可采用柚作砧木,红绵蜜柚和黄金蜜柚嫁接不可采用枳作砧木。     

高温对苹果花粉在花柱内萌发和生长的影响

以‘长富2号’红富士苹果为母本,‘红星’苹果为父本,研究35℃高温对苹果花粉在花柱内萌发和生长的影响。结果表明：温度影响花粉在柱头和花柱内的萌发及生长,与对照相比,高温促进花粉在柱头表面快速萌发,并加速花粉管在花柱内的伸长生长,但是在授粉72hN,高温处理下培养的花粉管形态出现花粉管变粗、弯曲并有瘤状小结的异化现象,同时,花柱发生褐变,并产生胼胝质,最终导致花粉管不能进入胚珠完成受精。     

一个柚新种质的亲缘关系鉴定

该研究以1个疑似柚新品种——‘迷你红柚’为实验材料,利用植物形态学观察、花粉电镜观察以及ISSR分子标记技术,以确定‘迷你红柚’与‘琯溪蜜柚’、‘邓肯葡萄柚’之间的关系。(1)从形态学观察可以看出:‘迷你红柚’果实的外形特征与‘邓肯葡萄柚’较相似,但与‘琯溪蜜柚’有较大的差别;而且‘迷你红柚’嫩叶的形态学特征与‘琯溪蜜柚’和‘邓肯葡萄柚’均有不同。(2)电镜观察发现:‘迷你红柚’花粉的萌发孔沟、花粉沟与‘琯溪蜜柚’和‘邓肯葡萄柚’相似,但花粉大小、外壁网眼与‘琯溪蜜柚’和‘邓肯葡萄柚’均有不同。(3)聚类分析结果表明,‘迷你红柚’和‘邓肯葡萄柚’遗传关系最近,但又有一定距离。研究认为,‘迷你红柚’不同于‘琯溪蜜柚’和‘邓肯葡萄柚’,极有可能是柚的一个新品种。     

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。     

柚子(Citrus grandis Osbeck)S-like核酸酶基因CgSL1的分离与特征分析

从柚子（Citrus grandis Osbeck)2个自交不亲和品种Guan溪蜜柚和度尾蜜柚的花柱中克隆出一个类似S核酸酶基因CgSL1（C.grandis S-likde RNase),它的cDNA序列全长1074bp,编码297个氨基酸,通过与其它植物S-like核酸酶和S核酸酶氨基酸序列进行比较,发现CgSL1类似于S-like核酸酶,与拟南芥中的RNS2一致性为62．5％。对CgSL1的表达分析表明该基因在花柱,花药,叶片不同器官以及花柱的不同发育阶段均有表达,且在花柱中的表达随衰老增强,由此推测它可能与衰老有关。     

十字花科10属种与油菜萝卜胞质不育系杂交的花粉萌发情况观察

对10属种十字花科植物与油菜萝卜胞质不育系杂交时花粉在柱头上粘合、萌发、花粉管伸长等情况进行观察。结果表明：（1）海甘蓝花粉粒粘合较难;（2）48 h内无瓣焊菜〖WTBX〗(Rorippa dubia)〖WTBX〗、毛果诸葛菜(Orychophragmus violaceus)、桂竹香(Cheiranthus cheiri)、海甘蓝(Crambe abyssinica)花粉管的伸长受阻于花粉萌发启动之时,花粉壁内形成胼胝质塞;播娘蒿、紫罗兰、荠菜花粉管伸长但未进入乳突细胞;芝麻菜花粉管进入乳突细胞而未进入柱头,‘浠水白’(Brassica campestris)、蓝花子有花粉管进入柱头及花柱而未进入胚囊。     

Guan溪蜜柚和度尾蜜柚自交和互交胚珠发育状况的比较

用石蜡切片技术对Guan溪蜜柚（Citrus grandis cv.Guanximiyou)和度尾蜜柚（C.grandiscv.Duweimiyou)自交和互交胚珠发育进行了比较观察。Guan蜜柚的自交授粉后胚珠发育有2种不同情形（Ⅰ型和Ⅱ型）,I型在授粉后5d的珠心组织细胞有变形现象。接着胚囊萎缩,但外珠被能生长发育到50d左右;Ⅱ型受精后胚乳发育异常,种子不能发育,度尾蜜柚自交胚囊中胚乳没有出现,在授粉后35d左右珠心组织和囊腔同时萎缩,互交胚珠发育正常。成熟果实有籽。表明两品种是自交不亲和的,导致了果实无籽。但两者的识别反应和阻抑部位不同,Guan溪蜜柚I型可能出现在受精前,Ⅱ型出现在受精后,而度尾蜜柚自交花粉管可能没有进入胚囊,是一种受精前的障碍。瘪籽的大小与不亲和障碍发生的时期有关,实验为进一步研究两品种的自交不亲和性遗传机理提供了实验证据。将有助于解决由于果实无籽导致的汁胞粒化,裂果,裂瓣等品质问题。     

琯溪蜜柚和度尾蜜柚自交和互交胚珠发育状况的比较

用石蜡切片技术对溪蜜柚 (Citrus grandis cv.Guanxim iyou)和度尾蜜柚 (C.grandis cv.Duweimiyou)自交和互交胚珠发育进行了比较观察。溪蜜柚的自交授粉后胚珠发育有 2种不同情形 (I型和 II型 ) , 型在授粉后5 d的珠心组织细胞有变形现象 ,接着胚囊萎缩 ,但外珠被能生长发育到 5 0 d左右 ; 型受精后胚乳发育异常 ,种子不能发育。度尾蜜柚自交胚囊中胚乳没有出现 ,在授粉后 35 d左右珠心组织和囊腔同时萎缩。互交胚珠发育正常 ,成熟果实有籽。表明两品种是自交不亲和的 ,导致了果实无籽。但两者的识别反应和阻抑部位不同 ,溪蜜柚 I型可能出现在受精前 , 型出现在受精后 ,而度尾蜜柚自交花粉管可能没有进入胚囊 ,是一种受精前的障碍。瘪籽的大小与不亲和障碍发生的时期有关。实验为进一步研究两品种的自交不亲和性遗传机理提供了实验证据 ,将有助于解决由于果实无籽导致的汁胞粒化、裂果、裂瓣等品质问题。     

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。     

中国樱桃与甜樱桃花粉原位萌发及花粉管生长的差异

以‘垂丝’、‘东塘’(中国樱桃)和‘莫利’、‘拉宾斯’(甜樱桃)为试材,分别于自花、异花授粉后不同时间切取花柱,用FAA固定,荧光染色后压片观察。结果显示,中国樱桃和甜樱桃的自花、异花花粉均能在柱头萌发,且其花粉管在花柱中表现为“极快—慢—快—稳定”的动态变化过程。但甜樱桃花粉萌发率高于中国樱桃,异花高于自花。中国樱桃自花、异花都有花粉管到达花柱基部;甜樱桃‘莫利’ב拉宾斯’和‘拉宾斯’自交有花粉管到达花柱基部,而‘莫利’自交的花粉管在花柱中上部已停止生长。结果表明,中国樱桃表现自交亲和性,而甜樱桃除人工诱变导致自交亲和的‘拉宾斯’等品种外,表现典型的植物配子体自交不亲和性。     

沙田柚花柱蛋白对花粉管生长的影响

本文以沙田柚及其授粉树种酸柚为材料,对其花粉在不同培养介质中的萌发率进行测定,筛选了０２～０４ｍｏｌ／Ｌ聚乙二醇－４００（ＰＥＧ－４００）＋００１％硼酸作为花粉萌发介质。用授粉后３ｄ的花柱提取液进行沙田柚、酸柚花粉萌发抑制实验,花柱提取液对自交花粉萌发无抑制作用,明显地抑制花粉管的生长。其中以花柱提取液的３５％饱和度硫酸铵级分对花粉管伸长的抑制最明显。模拟了花粉在雌蕊中的生长方式,进行花粉萌发生长,能收集到较多的花粉管,为研究花粉管的Ｓ－糖蛋白提供了材料。     

Polyamines and transglutaminase activity are involved in compatible and self-incompatible pollination of Citrus grandis

Pollination of pummelo (Citrus grandis L. Osbeck) pistils has been studied in planta by adding compatible and self-incompatible (SI) pollen to the stigma surface. The pollen germination has been monitored inside the pistil by fluorescent microscopy showing SI altered morphologies with irregular depositions of callose in the tube walls, and heavy callose depositions in enlarged tips. The polyamine (PA) content as free, perchloric acid (PCA)-soluble and -insoluble fractions and transglutaminase (TGase) activity have been analyzed in order to deepen their possible involvement in the progamic phase of plant reproduction. The conjugated PAs in PCA-soluble fraction were definitely higher than the free and the PCA-insoluble forms, in both compatible and SI pollinated pistils. In pistils, pollination caused an early decrease of free PAs and increase of the bound forms. The SI pollination, showed highest values of PCA-soluble and -insoluble PAs with a maximum in concomitance with the pollen tube arrest. As TGase mediates some of the effects of PAs by covalently binding them to proteins, its activity, never checked before in Citrus, was examined with two different assays. In addition, the presence of glutamyl-PAs confirmed the enzyme assay data and excluded the possibility of a misinterpretation. The SI pollination caused an increase in TGase activity, whereas the compatible pollination caused its decrease. Similarly to bound PAs, the glutamyl-PAs and the enzyme activity peaked in the SI pollinated pistils in concomitance with the observed block of the pollen tube growth, suggesting an involvement of TGase in SI response.     

Is Eucalyptus Cryptically Self-incompatible?

BACKGROUND AND AIMS: The probability that seeds will be fertilized from self- versus cross-pollen depends strongly on whether plants have self-incompatibility systems, and how these systems influence the fate of pollen tubes. METHODS: In this study of breeding systems in Eucalyptus urophylla and Eucalyptus grandis, epifluorescence microscopy was used to study pollen tube growth in styles following self- and cross-pollinations. KEY RESULTS: Pollen tubes from self-pollen took significantly longer than those from cross-pollen to grow to the base of the style in both E. urophylla (120 h vs. 96 h) and E. grandis (96 h vs. 72 h). In addition, both species exhibited reduced seed yields following self-pollination compared with cross-pollination. CONCLUSIONS: The present observations suggest that, in addition to a late-acting self-incompatibility barrier, cryptic self-incompatibility could be a mechanism responsible for the preferential out-crossing system in these two eucalypt species.     

Ethylene synthesis in petunia stigma tissues governs the growth of pollen tubes in progamic phase of fertilization

In the pollen-pistil system of petunia (Petunia hybrida L.) self-compatible and self-incompatible clones within 7 h after self-pollination, we determined the content of ACC (1-aminocyclopropane-1-carboxylic acid), the activity of two enzymes (ACC synthase and ACC oxidase), and the rate of ethylene production. Depending on the type of pollination, germination of pollen on the stigma surface and the pollen tube growth in the tissues of style were accompanied by different levels of ACC and ethylene release. The pollen-pistil system of the self-compatible clone contained twice more ACC than in the self-incompatible clone, whereas the pollen-pistil system in the self-incompatible clone produced 4–5 times more ethylene than in the self-compatible clone. For both types of pollination, ACC and ethylene were predominantly produced in the stigma tissues. The rate of ethylene production therein was 50 times greater than in the styles and ovaries, and the content of ACC was 100 times higher than in the styles and ovaries. Germination of male gametophyte after both types of pollination was accompanied by elevated ACC synthase activity (especially in the case of compatible pollination), whereas notable increase in ACC oxidase activity was manifested in growing pollen tubes after self-incompatible pollination     

AMONG- AND WITHIN-FLOWER COMPARISONS OF POLLEN TUBE GROWTH FOLLOWING SELF- AND CROSS-POLLINATIONS IN DIANTHUS CHINENSIS (CARYOPHYLLACEAE)

We compared the rate of pollen tube growth following self- and cross-pollinations both among and within flowers of two clones of Dianthus chinensis L. For among-flower comparisons, both styles of a flower were pollinated with either self- or cross-pollen. Within-flower comparisons were made between the two styles of the same flower, one of which was self-pollinated and the other cross-pollinated. Comparisons between flowers indicated that self-pollen grew slower than cross-pollen in both clones. However, differences in the growth rate of pollen tubes from self- and cross-pollinations were greater when comparisons were made between the two styles of the same flower than when pollinations were made in different flowers. These results suggest the existence of interstyle interactions in pollen tube growth.     

类整合素在烟草花粉萌发和花粉管生长中的作用

本文研究了动物整合素VnR抗血清及动物整合素专一性抑制剂含RGD的多肽对体外及半体内培养条件下烟草花粉萌发及花粉管生长的影响。结果表明在体外培养条件下,VnR抗血清及GRGDSP肽对花粉的萌发及花粉管的生长没有明显的抑制作用,但可抑制钙调素促进的花粉萌发和花粉管的生长;两者对柱头上进行的花粉萌发及在花柱里进行的花粉管生长也有一定程度的抑制。对类整合素在花粉萌发及花粉管生长中的作用进行了讨论。     

类整合素在烟草花粉萌发和花粉管生长中的作用

The effects of antiserum against human VnR integrin and integrin antagonist GRGDSP peptides on Nicotiana tabacum pollen germination and tube growth both in in vitro and in semi-vivo conditions were studied. No obvious inhibitory effects on pollen germination and tube growth in vitro were observed when anti-VnR serum or GRGDSP peptides was added to BK culture medium, but the enhancement of pollen germination and tube growth in vitro promoted by calmodulin was depressed by adding anti-VnR serum or GRGDSP peptides to BK culture medium. In addition to that, pollen germination and tube growth on stigma, as well as tube growth in styles were also inhibited at some extent by treating stigma and microinjecting GRGDSP peptides or anti-VnR serum into styles. The role of integrin-like proteins in regulation of pollen germination and tube growth in situ was discussed.     

烟草传粉前后生殖器官中细胞分裂素含量的变化

经免疫亲和层析系统纯化后,用间接竞争ＥＬＩＳＡ 法测定了烟草（Ｎｉｃｏｔｉａｎａ ｔａｂａｃｕｍ ）生殖器官在传粉前后细胞分裂素（ｔ－ＺＲ,ｉＰＡ）含量的变化． 开花前５ ｄ,花药和花丝中的细胞分裂素（ＣＴＫ）含量均达到最高值,以后随雄蕊发育逐渐下降． 授粉使花柱ＣＴＫ 含量急剧上升,授粉后１ ｄ 达到高峰,未经受粉的花柱ＣＴＫ 开花后下降．授粉后２ ｄ,子房中的ＣＴＫ 开始上升,在授粉后４ ｄ 达到最高值,而未受精的子房ＣＴＫ 含量开花后下降． 传粉后雌蕊中ＣＴＫ含量随花粉管生长而有规律地增加,ＣＴＫ 积极参与植物的传粉和受精过程     

Change in Cytokinin Levels in Reproductive Organs of Nicotiana tabacum Before and After Pollination

Immunoaffinity systems and indirect competitive ELISA were used to determine the changes of cytokinin (CTK) content in reproductive organs of Nicotiana tabacum before and after pollination. It was found that t-ZR and iPA existed in all reproductive organs tested, and their contents were higher in female than in male organs. On the 5th day before anthesis. CTK reached its peak value in anthers and filaments, then it declined. In styles, CTK content increased after pollination and reached its peak one day later. In ovaries, levels of CTK began to increase two days after pollination and reached its peak two days later, just at the time when fertilization took place. If unpollinated, CTK levels decreased both in styles and ovaries. Pollination and fertilization resulted in an increment of CTK in pistils, accompanied with the process of pollen germinfition and pollen tube growth. It could thus be inferred that t-ZR and iPA played some active role in pollination and fertilization.     

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.