60个苹果新品种S基因型鉴定

绝大多数苹果品种具有自交不亲和的特性，生产中需要搭配授粉树完成异花授粉。在苹果中已经分离鉴定了50多个S-RNase基因，对于一些新选育品种的S基因型至今未得到鉴定。利用S-RNase基因特异性PCR分析的方法鉴定了60份苹果新种质资源的S基因型，在60份苹果种质资源中，有56份品种的S基因型是首次鉴定到，并首次鉴定到了一个红肉苹果品种‘红色之爱’的S基因型，其S基因型是S19S？。新品种S基因型的鉴定能够为苹果育种亲本的选择及授粉树的合理搭配提供依据。     

八月酥等14个梨品种自交不亲和基因(S基因)型的鉴定

多数梨品种自花授粉不结实,生产上要合理配置授粉品种才能获得应有的产量。鉴定梨品种的S基因型,能为梨园合理配置授粉品种提供依据。以我国育成的梨品种为试材,利用特异引物PCR产物的聚丙烯酰胺凝胶电泳分离及DNA序列分析,鉴定出了14个梨品种的自交不亲和基因型(S基因型),并通过统计部分品种的田间人工授粉的坐果率以及应用荧光显微镜观察异花授粉后花柱内的花粉管生长情况来验证鉴定出的梨品种S基因型的可靠性。这些品种的S基因型为:八月酥S3S16、绿云S3S29、谢花甜S29S34、香茌S1S21、脆绿S3S4、新雅S4S34、雅青S4S34、伊犁红句句S22S28、猪嘴酥S19S22、假直把子S5S19、青魁S1S3、德胜香S3S29、张掖长把S3S29、金坠梨S21S34。     

‘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。     

新疆蔷薇科扁桃类植物自交不亲和性的分子机制

蔷薇科扁桃类植物是具有重要经济、研究价值的园艺果树类植物,该类果树植物家族均为自交不亲和性,自花授粉并不能正常结实,在其生长和生产过程中都需要进行异花授粉。扁桃类果树植物的自交不亲和属于配子型自交不亲和类型,是由花粉配子体核基因决定花粉与雄蕊的不亲和反应,即当花粉中的一个S等位基因与花柱中的一个等位基因相同时即表现不亲和。不亲和反应的细胞学特征是花粉管生长在花柱中受阻。该文对植物自交不亲和性方面的分子机制在蔷薇科扁桃类果树作物上的反映研究进展进行了综述,并着重阐述了新疆分布的独特的栽培扁桃和野生扁桃自交不亲和性S-位点基因的研究进展,以期为今后更深入的研究提供了重要的基础。     

果树自交不亲和花粉S基因研究进展

果树自交不亲和属于配子体自交不亲和,配子体自交不亲和至少由S位点的花柱S基因和花粉S基因2个基因决定,花柱S基因已被确定为S-RNase基因,而花粉S基因研究在近年才取得进展,一个在花粉中特异表达的基因SFB(Shaplotype-specificF-boxprotein)被发现,并且多方面的证据表明该基因是花粉S基因的最佳代表。就花粉S基因的筛选、该基因的结构特点、可能的作用机理等方面的研究进展予以综述。     

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。     

梨自交不亲和强度不同品种花柱S糖蛋白含量的差异

 提取梨( Pyrus) 自交不亲和强度不同品种花柱可溶性蛋白质, 应用等电聚焦电泳法在凝胶板上分离出S 糖蛋白, 用图像解析法进行定量。结果表明, 花柱总S 糖蛋白含量因品种而异, 自交不亲和强、弱及自交亲和的品种每1 􀀁g 花柱可溶性蛋白中含总S 糖蛋白分别为􀀂1. 3 ng 、0. 7 ~ 0. 9 ng、 0. 7 ng 。不同的S 基因所表达的S 糖蛋白量差异显著, 即使是同一S 基因所对应的S 糖蛋白量也因品种而异。     

11个延边地区栽培梨品种的S基因型鉴定

鉴定梨品种的S基因型可为合理配置授粉树、杂交亲本选配提供理论依据。为鉴定11个延边地区栽培梨品种的S基因型,利用梨的自交不亲和基因（S-RNase）特异引物FTQQYQ和anti-IIWPNV,对11个梨品种的基因组DNA进行S基因特异扩增,并对扩增片段进行回收、克隆、测序,使用生物信息学软件对各序列进行分析和同源性搜...     

'寒富'苹果自花结实性分析及S基因型鉴定

 本研究以苹果‘寒富’品种及其亲本为试材,研究其自交亲和性并鉴定其S基因型。田间授粉试验结果显示,‘寒富’品种表现为自花结实,其母本‘东光’品种亦自花结实;而父本‘富士’品种自花授粉结实率极低,初步推断‘寒富’品种的自花结实性可能遗传自其母本‘东光’。利用PCR技术鉴定'寒富'品种S基因型为S1S9。由于‘东光’品种存在同名异物问题,本研究除日本‘东光’品种外,对辽宁省果树研究所、吉林省果树研究所、沈阳农业大学等单位保存的中国'东光'品种的S基因型进行了鉴定,发现除辽宁省果树研究所保存的'东光'品种S基因型为S2S19,与‘寒富’品种无亲缘关系外,其它两个单位保存的'东光'品种中均存在S9基因型,可能为'寒富'杂交育种时的原始母本。     

‘奥嗄二十世纪’梨自交亲和性分子机制及遗传特性研究

 ‘奥嗄二十世纪’是自交不亲和性梨品种'二十世纪'的自交亲和性花柱突变体,花粉自交不亲和性功能正常,其自交亲和性突变的分子机制及遗传特性目前仍有争议。本研究通过田间自花及相互授粉以及基因组、mRNA转录和蛋白质水平比较,分析‘奥嗄二十世纪’、‘二十世纪’及其后代S-RNase基因的存在与否、表达特性及其在后代中的遗传。结果显示,‘奥嗄二十世纪’花柱S₂-RNase基因的核苷酸序列和表达特性与其原始品种‘二十世纪’的完全一样;而S4-RNase基因信号比其原始品种‘二十世纪’的弱,而且也在花柱中正常表达(包括转录和翻译水平),但表达量低;然而在其自交亲和后代基因组中检测不到S₄-RNase基因。本研究表明,‘奥嗄二十世纪’基因组中存在花柱S₄-RNase基因,但不能遗传给后代。     

秋子梨野生种和部分栽培品种交配亲和性研究

以秋子梨(Pyrus ussuriensis Maxim.)为试材,进行了花粉育性和交配亲和性研究。结果表明,参试的32份资源中有53%为雄性不育类型,可育品种间的花粉生活力存在明显差异,人工催花的花粉生活力低于自然开花,但对授粉结实无显著影响。南果梨、苹香梨、寒香梨等大部分栽培品种及山梨5个株系(野生秋子梨)都表现为自交不亲和。红花盖梨花期自交能够结实,但所结果实的种子不正常,为假亲和。龙香梨有一定自交结实力,花期人工自交花序坐果率达23.3%。部分具有自交不亲和性的梨品种蕾期自交授粉可提高结实率,但盛花后期授粉无显著影响。梨子代品种与亲代间互交,大部分组合表现亲和。南果梨×寒红梨组合交配亲和,但反交不亲和。梨同一类型不同品种间杂交结实正常,但亲本与其芽变品种间交配不亲和。     

Full compatibility is superior to semi-compatibility for fruit set in Japanese plum (Prunus salicina Lindl.) cultivars

Japanese plum belongs to the Rosaceae, which carry the S-RNase-based gametophytic self-incompatibility system. Therefore, these plum cultivars depend on cross-pollination for setting fruit. However, cultivar combinations may be genetically semi- or fully compatible. In apples and pears cultivated in Israel, where conditions for pollination are usually unfavorable, it was demonstrated that yields may be correlated with compatibility levels of pollinators. In the present study fruit-set of semi- and fully compatible plum couples was determined for a wide range of cultivars, in unfavorable and favorable conditions. Full compatibility was found superior to semi-compatibility, in both unfavorable and favorable conditions for pollination. These findings should be taken into consideration in determining the orchard designs of cultivars.     

Biochemical changes involved in self-incompatibility in two cultivars of olive (Olea europaea L.) during flower development

Biochemical analyses were carried out to characterise the basis of self-incompatibility in the olive (Olea europaea L.) cultivars ‘Amygdalolia’ and ‘Konservalia’. Multiple interactions between cells of different types, origin, and function occur in the pistil. Endogenous factors play important roles in ovary and fruit development before (Stage 1) and during pollination (Stage 2), and after fertilisation (Stage 3). Changes in carbohydrate, protein, H₂O_2, and calcium ion concentrations, and in peroxidase, catalase, superoxide dismutase, and ascorbate peroxidase activities in pistil tissue before and during pollination, and after fertilisation were investigated. In both cultivars, H₂O₂ concentrations were significantly higher in pistil tissue before pollination, after which they started to decrease in Stage 2 and continued non-significantly in Stage 3. Peroxidase, catalase, superoxide dismutase, and ascorbate peroxidase activities were lower at Stage 1 and Stage 2, whereas these enzyme activities increased at Stage 3 in both cultivars. The lowest concentration of calcium ions was observed at Stage 1, whereas at Stage 2, calcium ion concentrations increased and reached their highest level, then decreased at Stage 3. Calcium ion concentrations in ‘Konservalia’ were higher than in ‘Amygdalolia’. In ‘Konservalia’, the highest protein concentration was observed at Stage 2. No significant differences were found in carbohydrate concentrations between the two cultivars. The biological significance of the presence of these products may differ between Stage 1, when they have a defence function, Stage 2 when there are interactions between pollen and pistil, and Stage 3 after fertilisation. This study provides support for the hypothesis that there is a correlation between self-incompatibility, stress-related enzyme activities, and calcium ion concentrations in the pistils of olive.     

Physiological Breakdown in Stored Monarch Plums

Summary

Several pollination studies carried out on different, self-incompatible almond cultivars and seedlings have shown the presence of variable levels of fruit set following self-pollination that could be attributed to partial self-incompatibility (PSI). PSI is an intermediate reproductive behaviour, described in some angiosperm species, which indicates that self-incompatibility is a quantitative and plastic trait. The present study was performed to substantiate the occurrence of PSI in almond by studying ten cultivars that are traditionally considered to be self-incompatible (four of which had previously shown fruit set after self-pollination). These cultivars were analysed by microscopic observations of the progression of pollen tubes through the pistil following controlled self-pollination, determinations of fruit set after bagging flower buds, and molecular identification of the parentage of the fruits obtained using consensus PCR-primers for Prunus S-RNase alleles. The results showed that, in nearly all cases, the pollen tubes did not enter the ovary and, from a total of 5,349 bagged flower buds, only 17 fruits were obtained. In all cases, PCR analysis of the plants obtained after germination of these seeds showed a band corresponding to an S-RNase allele not present in the maternal progenitor, clearly indicating that they could only have arisen from cross-pollination.Therefore, the low fruit set values observed in some of the cultivars studied were not due to a breakdown in the self-incompatibility response that confers PSI, but to very low rates of contamination with foreign pollen. These results corroborate the self-incompatibility phenotype of the cultivars studied here, and highlight the importance of ascertaining the identity of the parentage in any progeny obtained after bagging.     

不同发育阶段甜樱桃花授粉后花粉萌发和花粉管生长的差异

选用自交不亲和性程度不同的2个甜樱桃品种莫利和拉宾斯为试材,分别在开花前、开花当天和开花后进行莫利自花、拉宾斯自花和莫利×拉宾斯异花授粉,并于授粉后12、24、48、72、96h切取花柱,用FAA固定,荧光染色后压片观察。结果表明,3个组合的花粉萌发率都以开花当天授粉的最佳,拉宾斯自交和莫利×拉宾斯的花粉管生长状况也以开花当天授粉的最佳,但莫利自交的花粉管生长状况却以花前第4天最佳,花后第4天次之,而当天授粉的最差。因此认为,甜樱桃自交不亲和程度与其雌蕊的发育状态密切相关,蕾期授粉和延迟授粉都能够在一定程度上克服甜樱桃的自交不亲和性,但蕾期授粉的作用更加明显。     

甜樱桃S基因型PCR鉴定技术研究

绝大部分甜樱桃品种自交不亲和,因此自交不亲和基因型的鉴定对于生产具有重要的意义。以甜樱桃主栽品种为试材,建立基于PCR技术的甜樱桃品种S基因型鉴定技术。试验根据已发表的樱桃S基因序列设计了2对引物组合BFP73/74,BFP93/94,结合S1、S5基因的特异扩增引物,利用扩增片段长度的不同,就可以对樱桃品种的S基因型进行鉴定。通过对已知S基因型品种基因组的扩增,最终建立了基于PCR技术的甜樱桃品种S基因型鉴定技术。     

Study on Fruitset and Pollen-Compatibility Status in Sweet Cherry (Prunus avuim L.) Cultivars

Prunus avium L. is one of the most important temperate zone fruits in the world. Most of the cherry cultivars always have difficulties of fertilization and fruit set due to self-incompatibility, so they need suitable and compatible pollinizers for commercial fruit production. In this experiment, pollination compatibility of cherry cultivars of ‘Napoleon’, ‘Burlat’, ‘Zhan’, and ‘Lambert’ was studied as both pollen recipients and donors. To determine the compatibility or incompatibility of pollinizers, percent of fruit set was calculated. This research was carried out as a factorial experiment in a randomized complete block design. The final average fruit set in studied cultivars was different under self or cross-pollination. The results showed that ‘Napoleon’, ‘Zhan’ and ‘Lambert’ cultivars are the suitable pollinizers for ‘Burlat’. Also, ‘Burlat’ is a cultivar which produced many fruits under self-pollination. Therefore, ‘Burlat’ can be used as monoculture for orchard establishment. Results showed that cultivar ‘Napoleon’ was cross-incompatible with ‘Lambert’. ‘Zhan’, ‘Napoleon’ and ‘Lambert’ cultivars are founded self-incompatible and require cross-pollinations to obtain fruits.

     

苹果新品种自交不亲和基因型的分子生物学鉴定

根据保守氨基酸序列FTQQYQ 和anti-¹/_M IWPNV 设计苹果自交不亲和基因引物P1 : 5′2 TT2TACGCAGCAATATCAG23′; P2 : 5′2 ACGTTCGGCCAAATA /CATT23′, 利用PCR - RFLP分析方法, 以河北省新引进的苹果品种美国8号、昂林、松本锦、华冠、南方脆幼叶为试验材料, 鉴定其自交不亲和基因型分别为S₉S₂₇、S₁S₉、S₉S ₉、S₂S₉、S₂S₉。     

中国原产果梅自交亲和变异品种花粉决定基因SFB的插入突变

 果梅是配子体自交不亲和树种,但在长期进化过程中也出现了自交亲和品种。以果梅典型的自交不亲和品种‘南高’和自交亲和品种‘甲州小梅’为对照,采用田间自交授粉试验对原产于中国的‘四川白梅’和‘长农17’两个果梅品种进行了自交亲和性鉴定,结果表明：其自花授粉坐果率分别为30.73%和16.27%,属于自交亲和品种。进一步通过AS-PCR分析发现花粉SFB基因存在一段插入序列,造成了分子水平的基因突变。推测该基因的突变使‘四川白梅’和‘长农17’SFB功能发生改变,从而自交亲和。     

Effects of putrescine application on fruit set in ‘Housui’ Japanese pear (Pyrus pyrifolia Nakai)

In Japan, artificial pollination is practiced to improve fruit set as well as fruit size and to obtain perfect round shapes in ‘Housui’ (S₃S₅) Japanese pear and most commercial cultivars that present gametophytic self-incompatibility. The period of artificial pollination application is limited by environmental conditions and the availability of a labor force. The present study aimed to determine if pre-pollination applications of putrescine (1.0 and 0.01 mM) positively affect fruit set in ‘Housui’ after hand-pollination. During 2002 and 2003, fruit set in late pollinated flowers increased with pre-pollination putrescine applications and concentrations of 1.0 mM had a better effect than 0.01 mM. In vitro pollination tests revealed that higher pollen germination was present in the stigmas of flowers sprayed with 1.0 mM of putrescine compared to the controls. Putrescine application did not affect post-pollination ethylene production in the styles. Moreover, in the styles of non-treated flowers, an increment of putrescine content and ethylene production after pollination was found, suggesting post-pollination responses and confirming the lack of antagonism between polyamines and ethylene at least during this process.