苹果梨树最佳授粉受精时间观测

苹果梨是自花不结实品种。苹果梨在开花后5d（天）内均有很高的授粉能力，其中，花后2－5d（天）坐果率最高，花朵坐昆率为90.0%－96.7%，花后第6d（天）授粉能力开始降低，第7d（天）花朵坐果率已降至10.0%，第8d（天）失去授粉能力。苹果梨授粉10h（小时）后花粉管开始进入花柱，24h（小时）后花粉管大部分已进入花柱，48h（小时）到达子房中部，56h（小时）后开始进入子房，96h（小时）绝大部分花粉管到达子房完成受精过程。     

建阳桔柚花粉育性、授粉受精和胚胎早期发育的观察

以3年生枳砧建阳桔柚为试材,观察花器官结构、花粉育性、授粉后花粉管生长过程和早期胚胎发育情况。结果表明,建阳桔柚的花为完全花,花粉染色活力为66.42%,花粉萌发率为8.68%。自花和异花(沃柑花粉)授粉4h后花粉都开始萌发;授粉8h后异花授粉的花粉大量萌发,自花授粉萌发量比授粉4h时多,但低于异花授粉;异花授粉24h后少量花粉管伸入子房,异花授粉48h后大量花粉管进入子房中且部分完成受精;自花授粉48h后花粉管进入子房,但并没有受精成功,自花授粉72h后可观察到少量受精成功。自花授粉于授粉2周后观察到胚珠内部出现空腔、早期败育;异花授粉于授粉3周后观察到部分胚珠内部出现空腔、早期败育。     

果梅花粉原位萌发及花粉管生长特性的研究

以软条红梅为试材,分别于白花授粉和异花授粉后的1、3、6、12、24、48、72、96、120 h切取花柱,用FAA固定液处理、荧光染色后压片观察,结果表明:异花和自花花粉都能在柱头上萌发,但在授粉后的48 h之内异花花粉的萌发率显著高于自花花粉的萌发率,如在授粉6 h后,异花花粉萌发率高达58％,而自花花粉的萌发率仅为23％。花柱内异花花粉管生长速度明显比自花的快,在授粉后花粉管到达花柱上部、中部及基部的数目均是异花高于白花。如授粉24 h后异花花粉管平均有30余根穿过花柱上部,约有14根到达中部,有2根到达基部;而自花授粉的花粉管分别仅有18、7和0根,且白花授粉时在72 h内均未见有花粉管到达花柱基部,主要在花柱中部前后停止生长。结果表明软条红梅自花授粉的花粉管主要是在花柱中部被抑制而停止生长,从而不能完成受精,表现出典型的配子体型自交不亲和性。     

苹果自花授粉的花粉管结构变化及S-RNase定位

 苹果‘红星’品种授粉处理24 h后, 自花授粉比异花授粉的花粉管生长缓慢。授粉处理48 h后, 可观察到自花授粉花柱通道组织的花粉管内有二裂粒子增加、管壁破裂和管内物质溢出、外层管壁增厚等现象。苹果气球状花期未授粉的花柱及自花和异花授粉24 h后且无花粉管存在的花柱切片显示, S-RNase位于花柱通道组织的细胞内。自花和异花授粉48 h后有花粉管通过的花柱切片显示, S-RNase位于花柱通道组织的细胞间隙和花粉管内。     

甜樱桃花粉原位萌发荧光显微镜观察

以"美早""早大果"和"斯坦拉"甜樱桃为试材,分别于自花、异花授粉后不同时间(27、51、75、99h)切取花柱,通过荧光压片法观察各授粉组合的花粉原位萌发及花粉管生长情况,以期为研究甜樱桃授粉机理提供依据。结果表明:3个甜樱桃品种的花粉均能在自花或异花柱头上萌发,且于授粉27h后趋于稳定,不同授粉组合间花粉萌发量只在数量上表现出差异,其结果并不会影响坐果率。在花粉管生长情况上,不同授粉组合表现出显著差异。授粉后51~99h,亲和花粉管不断向花柱基部延伸,进入子房;而不亲和花粉管在花柱中上部受到抑制,停止生长。同为自花授粉,"斯坦拉"自交的花粉管能够到达花柱基部,表现为自交亲和;而"美早"和"早大果"自交则表现为极强的不亲和性。异花授粉中,"斯坦拉"与"美早"和"早大果"均表现为亲和性较好。综上所述,"斯坦拉"无论是自花还是异花授粉,花粉管均能到达花柱基部,完成受精,是较为理想的授粉树。而"美早"和"早大果"只有在异花授粉时花粉管才能到达花柱基部,而自花授粉的花粉管在花柱中上部已停止生长,故在实际生产中需配置授粉树。     

外源多胺对鸭梨自花授粉花粉管生长及自交坐果的影响

以鸭梨为试材,进行不同浓度外源多胺处理,通过离体花粉培养,荧光显微动态观察和花序、花朵坐果率调查,研究了外源多胺处理对鸭梨自花授粉花粉管生长及自交坐果的影响,以期为外源多胺调控鸭梨自交不亲和性提供重要依据。结果表明:适宜浓度的外源腐胺(Put)、亚精胺(Spd)、精胺(Spm)处理后的鸭梨花柱提取液,能够提高花粉萌发率和促进花粉管生长,最适宜的Put浓度和Spd浓度为0.250 mmol·L~(-1),Spm浓度为0.050mmol·L~(-1)。鸭梨自花授粉72h后,花粉管先端膨大,在花柱中部停止生长。3种多胺处理均能使其穿过花柱中部到达基部,其中以0.250mmol·L~(-1) Spd处理花粉管到达基部最多。不同浓度外源多胺均能提高鸭梨自花授粉花序坐果率和花朵坐果率,其中以0.250mmol·L~(-1) Put表现最佳。     

梨不同花龄自花与异花授粉的花柱内花粉管生长及坐果率的比较

对自交不亲和强度不同的梨品种丰水和菊水的不同花龄花进行自花授粉和丰水×菊水异花授粉后,用荧光显微镜观察花粉萌发及生长特性。结果表明,开花当天及以后的自花和异花授粉花粉在柱头上均有较高的萌发率;自花和异花花粉管在花柱内的生长速度及停止生长的位置不同,丰水自花授粉后花粉管生长的短且慢,菊水自花花粉管生长的长且快,而异花授粉的花粉管生长最快。自花授粉后,不同花龄对花粉管生长影响表现为:开花4d前的蕾期授粉以及开花4d后的延迟授粉,花粉管生长长度显著的长。开花6d前的各种授粉组合,花粉管生长均较差。这与田间授粉坐果率的高低基本一致,即整体表现为各个花龄的授粉都以异花授粉坐果率最高,其次为菊水自花授粉,丰水自花授粉坐果率最低。另外,自花授粉中,蕾期授粉和延迟授粉(开花2d后除外)坐果率都有提高,而异花授粉中蕾期授粉和延迟授粉的坐果率却有所下降。     

9个新疆杏品种间杂交亲和性研究

【目的】了解新疆杏品种间杂交亲和性,为授粉树的配置提供理论依据。【方法】以9个新疆杏品种为试材,测定各品种的花粉量、花粉活力,设置不同品种间72个授粉组合,测定杂交组合的花粉管生长状况及坐果率等指标,以判定不同杂交组合的亲和性。【结果】9个品种平均花粉量为每花药1 474.4粒,品种间差异极大。各品种平均花粉萌发率为44.0%,其中,‘库曼提’花粉萌发率较高,为80.0%,‘树上干’较小,仅17.5%。不同授粉组合的平均坐果率为25.9%,较自然授粉平均高2.7倍,以‘阿克阿依’为母本,‘轮台白杏’为父本的授粉组合坐果率最高,达64.2%。荧光显微观察结果表明,所有杂交组合花粉多在授粉后24 h萌发并伸长,授粉96 h后到达花柱基部进入子房,但各组合花粉在花柱原位萌发的速度及花粉管生长速度存在差异,以‘佳娜丽’为母本,‘库尔勒托永’为父本等9个组合,花粉管生长延长到花柱中下部后,停止向下生长,只有个别能正常生长到达子房。【结论】各品种间花粉量、花粉活力差异极大。在72个授粉组合中,坐果率小于6%的授粉组合,花粉管未到达花柱基部或到达基部的速度慢,比率小,表现为杂交不亲和;坐果率较高的授粉组合,花粉管到达花柱基部的速度快,比率大,亲和性强,适宜搭配建园。     

自花和异花授粉对蓝莓花粉管动力学的影响

以‘园蓝’和‘蓝金’为试材,以‘灿烂’为异花授粉品种,开展自花和异花授粉,授粉后不同时间取样观察花粉原位萌发和花粉管生长情况,分析自花和异花授粉对参试品种花粉管动力学的影响。结果表明:自花和异花授粉对‘园蓝’的花粉管动力学有显著影响,其自花授粉花粉管生长速度显著低于异花授粉,胚珠的退化速度显著加快,至授粉后120h,异花授粉处理被花粉管生长穿过的花柱比率显著高于自花授粉,表现出自交不亲和性;自花和异花授粉对‘蓝金’的花粉管动力学有一定影响,但至授粉后120h,其异花授粉处理被花粉管生长穿过的花柱比率与自花授粉无差异,表现为自交亲和。综合分析表明,参试材料自花授粉受精生物学特性具有明显的种群特异性,‘园蓝’自交不亲和反应发生在花柱基部,属于配子体自交不亲和中的合子前不亲和类型。     

密斯蒂蓝莓花粉形态及授粉亲和性观察

以密斯蒂蓝莓为试材,进行花粉粒电镜扫描及自花授粉和异花授粉试验,观察花粉管萌发及生长动态荧光显微,调查不同授粉方式的着果率,测定果实品质。结果表明,密斯蒂花粉形态均为四合花粉,呈四面体排列,近似圆角三角形,极轴×赤道轴为28.5μm×26.02μm;荧光显微观察发现自花授粉和异花授粉8h花粉开始在柱头萌发,14h有大量花粉萌发,花粉管进入花柱;24h花粉管进到花柱底部;48h花粉管进入胚珠,到达胚囊,完成双受精,自花授粉和异花授粉均有较好的亲和性;密斯蒂异花授粉着果率最高,两种授粉方式果实品质无显著性差异。     

杏品种授粉生物学研究

以红丰、新世纪等11个杏品种及特新一号等4个新品系为试材,对自交与杂交亲和性、花粉管形态及花粉、花柱氨基酸组分进行了研究,旨在为杏生产和进一步研究提供科学依据。结果表明:(1)按照白花授粉坐果率≥6％为自交亲和这个标准,红丰、新世纪等10个品种为自交不亲和品种(SI),而特新一号等4个新品系是自交亲和的(SC);(2)选自同一杂交组合不同姊妹系的红丰与新世纪杏品种相互授粉坐果率仅为0~0.73％,属于杂交不亲和;(3)不同品种间相互授粉的坐果率差异很大,据此筛选出特早红、红玉杏及凯特等品种是红丰杏适宜的授粉品种;而新世纪用骆驼黄及金太阳等品种授粉的坐果率最高;(4)自交不亲和组合在授粉后72 h花粉管到达花柱3／4处,末端膨大,停止生长,而自交亲和组合的花粉管可以继续生长,并在授粉后96 h进入子房;(5)自交不亲和品种(系)与自交亲和品种(系)花粉的17种氨基酸含量并无显著差异,而花柱氨基酸的组分与花粉氨基酸组分有所不同,其中自交不亲和品种(系)酪氨酸、苏氨酸、甲硫氨酸和精氨酸含量明显高于自交亲和品种(系),差异显著或极显著。     

Pollen tube growth in styles of apple and almond flowers after spraying with pesticides

Summary

The timing of fungicide and antibiotic applications in fruit crops often overlaps flowering and pollination. Numerous studies report detrimental effects of chemical applications on pollination, fruit set and yield. However, field results can be quite variable even when similar compounds are tested on the same crop. Problematic is that field variables unrelated to the effects of pesticides on reproductive activities can mask results. In the current study, we specifically investigated the effects of selected pesticide sprays applied to stigmas, on subsequent pollen germination and tube growth. Using two model systems, apple and almond, sprays were applied to detached emasculated flowers under constant laboratory conditions using an electronically controlled robotic apparatus which simulated a field sprayer. flowers were pollinated at specific times after spraying, and pollen tube numbers and growth assessed using fluorescence microscopy. The pesticides evaluated were for apple: captan, myclobutanil and streptomycin; and for almond: azoxystrobin, myclobutanil, iprodione and cyprodinil. In apple, captan significantly reduced pollen germination, by 20% compared with water controls when pollination was conducted 18 h after treatment. The number of tubes that reached the basal part of the style within 20 h was not affected. In contrast, myclobutanil and streptomycin had no significant effect on pollen tube growth. With almond, none of the sprays significantly affected pollen germination or tube growth.     

酥梨受精过程的观察

选用中华玉梨、新水、红香酥、早酥4个品种的花粉给酥梨授粉,授粉后8、24、48、72、96h分别取样,然后进行固定、解离、染色等处理,制成压片和切片进行受精过程的观察,旨在探索酥梨授粉受精内在规律,为生产上促进酥梨授粉受精及提高坐果提供理论依据。结果表明:各授粉品种授粉后8h,花粉管即进入花柱,24h花粉管已布满子房组织中;酥梨以合点受精为主;各授粉品种与酥梨亲和力良好,完成受精时间为:中华玉梨48h、新水48h、红香酥72h、早酥72h,花后2周其花朵坐果率可达74.49%～87.26%。酥梨在花期温度不低于3.3℃的情况下,有利其完成授粉受精及坐果过程。     

Rain effect on pollen–stigma adhesion and fertilization in almond

The fact that rainy weather negatively affects production in almond is well-known by growers. However, as far as we know, no previous study has been carried out in this species to determine if the rain is able to wash off pollen grains from the stigma surface and consequently affect fertilization efficiency. To answer this question, flowers of the self-compatible almond cultivar ‘Antoñeta’ were emasculated on branches of trees in the field, pollinated by hand with pollen of the cultivar ‘Marcona’, and sprayed with water to simulate the rain effect at 4, 8, 24 and 48 h after pollination. A control for which no treatment was applied was also included in the assay. Seven days after spraying the flowers, the number of germinated pollen grains on the stigma was recorded in 10 pistils for each treatment by fluorescent microscopic observation. In addition, for each treatment the initial and final fruit set were determined at 30 and 60 days after pollination, respectively. The following year additional assays were performed in the field and in the laboratory in order to complete the study with earlier treatment times. Thus, an assay of rain simulation in the field at 0 and 2 h, and other consisting on immersing a branch with flowers and shaking it in a water container in the laboratory at 0, 30, 60 and 120 min after pollination were performed. One sample more was immersed, shaken, then pollinated and again immersed and shaken. The results from field assays showed that adhesion of pollen grains to the stigma was very quick and strong, and the treatments did not significantly affect pollen–stigma adhesion and subsequent fruit set. On the other hand, laboratory results showed that all washing treatments decreased the number of germinated pollen grains on the stigma, mainly when the flowers were immersed before pollination. In conclusion, the results showed that the simulated rain was not able to completely wash off pollen grains from the stigma surface, although it seems to affect adhesion in forthcoming pollinations.     

杏(Prunus armeniaca)自交不亲和强度及其授粉受精相关特性

为了解杏自交不亲和性强度与授粉受精相关特性的关系,以自交亲和(Self-compatibility:SC)品种凯特(Prunus armeniaca L．cv．Katy)和自交不亲和(Self-incompatibility:SI)品种新世纪(P.armeniaca L.cv．Xinshiji)及凯特×新世纪杂种群体为试材,荧光显微镜观察自交亲和与自交不亲和杏花粉管生长动态。结果表明,授粉后初期,自交亲和性与自交不亲和性的杏花粉都能正常萌发、生长,但是在花粉管生长延伸到花柱1／2以后,自交亲和性的花粉管能顺利进入子房,而自交不亲和性的花粉管多数顶端膨大呈球形,停止向下生长,只有极个别能正常生长到达子房;杂种后代的可溶性蛋白含量和RNA酶比活力,与亲本相比无明显的趋中变异表现,而且在自交亲和与自交不亲和杏之间无显著性差异。     

Mentor pollen effect on the in-vivo germination of self-incompatible apple pollen

Investigations were carried out to ascertain to what extent “mentor pollen” can influence the performance of self pollen in apple styles. Self-incompatible ‘Starkrimson’ flowers on branches, placed at room temperature or in a growth chamber, were (1) self-pollinated, (2) cross-pollinated with compatible untreated or (3) irradiated (150 krad) ‘Golden Delicious’ pollen, or (4) pollinated with a mixture (2:1 w/w) of irradiated (mentor) pollen and self pollen. After 18 h, nearly the same proportion (> 90%) of the self-incompatible pollen had germinated on the stigma and penetrated the apical part of the style as compatible pollen. However, subsequent growth of the self-pollen tubes was inhibited. After 48 h, less than 1% had reached the stylar base as compared with 21–24% of the compatible pollen tubes. Irradiation strongly impaired pollen vitality: only 1% of the tubes had reached the base of the style after 48 h. The mixture of the irradiated mentor pollen and self pollen performed better. In this case, 6.4–9.9% of the tubes was observed in the basal part of the style, inferring that tube growth of the self pollen was significantly promoted by the mentor pollen.     

Effective pollination period in durian (Durio zibethinus Murr.) and the factors regulating it

The effective pollination period (EPP) of durian was determined by both delayed and bud pollination, during which reproductive factors affecting the EPP, e.g., stigma receptivity, pollen tube growth in the style, and ovule longevity were studied histologically. This study was conducted in three distinct locations in Thailand, namely, the Chantaburi Horticultural Research Center and two private orchards in Chantaburi and Trat provinces. Results from artificial pollination revealed that at anthesis, the durian flower is receptive and has a high fruit set ratio. A mean fruit set of 50% was obtained at anthesis in the private orchard in Chantaburi province. However, the EPP of durian was found to be very short, lasting for only one night; the fruit set from pollination on the morning after anthesis ranged from 0% to 3.4%. No fruit set occurred following pollination 24 or more hours after anthesis. When compared with the flowers of other fruit species, the durian flower has a unique feature in that it blooms overnight; the following morning, there is abscission of all parts of the flower, except the gynoecium. Thus, EPP appears to be synchronized with flower longevity. On the other hand, the durian flower was receptive several hours before anthesis. The results of chemical tests, including the hydrogen peroxide test and Perex-Test^®, for the evaluation of stigma receptivity appeared to be in agreement with the EPP. However, fluorescent microscopy revealed that pollen could germinate even in the stigmas pollinated 48 h after anthesis, but the number of pollen tubes at the top of the style rapidly decreased from 34.6 (at anthesis) to 0.5 (48 h after anthesis). A correlation test demonstrated a higher correlation coefficient between the fruit set and the number of pollen than that between the fruit set and the result of Perex-Test^®. This indicated that pollen tube penetration or elongation in the style was inhibited, probably due to the deterioration of nutritional support from the pistil to the pollen tubes; this can be a limiting factor of the EPP in durian.     

‘金坠梨’自交亲和性突变机制的初步研究

 自交不亲和性品种鸭梨的芽变品种‘金坠梨’自花授粉能结实, 但其自交亲和性突变机制至今不祥。本研究结果表明, 鸭梨自花授粉结实率仅2% , 自花花粉管在花柱中上部已经停止生长, 而金坠梨自花授粉结实率高达76% , 自花的花粉管能正常生长至花柱基部; 鸭梨花柱能够接受金坠梨花粉并受精结实, 而金坠梨花柱却与鸭梨花粉杂交不亲和; 进一步鉴定出了鸭梨和金坠梨基于S-RNase基因的S基因型, 发现两者S基因型相同, 均为S₂₁S₃₄ ; 通过克隆两品种S-RNase基因cDNA全长序列并进行比较分析发现, 该两品种的1对雌蕊S-RNase基因在核苷酸序列上完全相同, 且S-RNase基因均正常表达, 表达量没有明显差异, 从分子水平上证实了金坠梨在花柱方面和鸭梨并无差异, 可能是花粉S 基因发生突变, 导致了自交不亲和性功能的丧失, 表现出自花授粉能够结实。     

Effect of pistil age and pollen parent on pollen tube growth and fruit production of pistachio

Female pistachio (Pistacia vera) trees were hand pollinated with pollen from three different male genotypes 0,1,2,3 and 4 d after anthesis. Results were measured by mature fruit set and by fluorescence microscopy of pollen germination and tube growth in the pistil. Maximum pollen tube growth and fruit set of split nuts was achieved following pollination within two days of anthesis. Three day old pistils supported pollen tube growth, but fruit set was low. Four day old pistils supported little pollen tube growth or fruit set. Significant differences were also apparent between pollen parents, with low pollen tube growth and fruit set following interspecific pollination with P. atlantica pollen. All pollen tubes were observed to penetrate chalazogamously, and parthenocarpic production of blank fruits occurred in both unpollinated and pollinated treatments. The results indicate that for optimum fruit production of pistachio, pollen transfer must be achieved within 2 d of anthesis and pollination by P. atlantica should be avoided.     

日光温室条件下甜樱桃授粉受精期间环境因子对花粉行为的影响

以甜樱桃(Prunus avium L.)红灯为试材,采用荧光显微技术研究了日光温室和露地条件下甜樱桃花粉行为的差异,并观察记载了授粉受精期间日光温室和露地环境因子的变化情况。结果表明,环境因子影响了甜樱桃的花粉行为,日光温室内花粉萌发和花粉管伸长速率明显落后于露地。日光温室条件下,授粉后6h花粉萌发,120h花粉管进入子房;而露地条件下,授粉后3h花粉萌发,72h花粉管进入子房。其中,气温是影响日光温室甜樱桃授粉受精进程延缓的主要环境因子。