Incompatibility in apricot (Prunus armeniaca L.): Methodological considerations

Summary

The recent introduction of new self-incompatible varieties of apricot has required further research on the extent of self-incompatibility in apricot and its genetic determinism. The different laboratory and field techniques utilized to date for incompatibility studies in apricot have often given markedly diverging results, leading to difficulty of interpretation. The purpose of this study was therefore to make a critical assessment of the reliability of the various methods, to obtain a more precise tool for compatibility studies. Forty-eight apricot crosses were assessed over a two-year period and the results compared using the normal field techniques (active pollination without bagging, active pollination with bagging and passive pollination) and laboratory tests (fluorescence test). The present study reports on some aspects of the different techniques and offers methodological suggestions for improved practical application of the various tests.     

Cold and heat requirements of the apricot tree (Prunus armeniaca L.)

Summary

The cold units (CU) and heat units (HU) required for the flowering of 13 apricot cultivars (Prunus armeniaca L.) cultivated in AbaraÂn (Murcia) have been established. The end of the dormancy period was determined on the basis of the dry-weight increase of the flower buds. The tests in which the dry weight was measured after forcing the flower buds at 208C turned out to be more precise in detecting the beginning of the reactivation phase than those in which they were not forced.     

Development of SCAR markers to differentiate between mume (Prunus mume Sieb. et Zucc.) and apricot (P. armeniaca L.)

Summary

Mume (Prunus mume Sieb. et Zucc.) and apricot (P. armeniaca L.) are similar in fruit and tree morphology, and exhibit high cross- and graft-compatibility with each other. It is therefore difficult to differentiate mume and apricot cultivars on the basis of morphological and phenotypical characteristics. Molecular markers were developed to differentiate nine mume from ten apricot cultivars. Four dominant, random amplified polymorphic DNA (RAPD) markers that can discriminate between mume and apricot cultivars (designated OPA15₆₂₈, OPO10₅₅₀, OPO20₂₅₉, and OPU03₄₁₅) were identified from 21 decamer primers. Two RAPD markers (OPO10₅₅₀ and OPU03₄₁₅) were developed into dominant sequence-characterised amplified region (SCAR) markers (SCO10 and SCU03). These SCAR markers could differentiate between all mume and apricot cultivars.     

杏的脱毒及组培快繁研究

杏(Prunus arm eniaca L.)作为主要核果类果树树种之一,在世界各大洲都有栽培。随着研究的深入,研究人员发现,给杏树生产带来严重危害的是病毒,并且已应用ELISA方法检测出杏树病毒有李子苦痘病毒(PPV)、李枯叶坏死坏斑病毒(PNRSV)、李矮缩病毒(PDV)、苹果花叶病毒(ApM V)、苹果     

普通杏品种SSR遗传多样性分析

利用来自杏、桃和扁桃的25对SSR多态性引物对66个普通杏品种进行了PCR扩增及变性聚丙烯酰胺凝胶电泳,共检测出284个等位位点,每对引物的等位位点数在3～17之间,平均为11.36。通过NTSYS软件计算得到的Dice相似性系数为0.083～0.987,平均值为0.370,表明中国普通杏种质资源的遗传多样性丰富。UPGMA法聚类将66份材料分为5个组。SSR标记反映出的品种间亲缘关系与根据地理生态类型对杏种质的分类结果基本一致。来自四川和贵州的多数品种独立聚成一组,表现出与其它品种较远的亲缘关系,该地区可能存在特异性较高的种质资源。证实了扁桃基因组SSR引物可用在杏SSR标记的研究中。     

不同引物组合对中国杏品种资源S基因特异扩增效果比较

 利用已报道的5对蔷薇科李属通用引物组合对起源于我国的30个杏(Prunus armeniaca L.)品种进行了S等位基因的专一性PCR扩增（S-allele specific PCR），并依据扩增系数和扩增成功率两个指标对扩增效果进行了评价。结果表明：①不同的引物组合对参试杏品种的扩增系数和扩增成功率差异很大，其中引物组合EM PC2consFD+EM PC3consRD扩增系数和扩增成功率均为最高（66.7%和53.3％），效果最好；②引物组合EM PC2consFD+EM PC3consRD对‘华县大接杏’和‘猪皮水杏’未能扩增出任何条带，而PruC2+PruC4R和PaConsⅡ F+PaConsⅡ R两对组合却能对其扩增出两条带；此外，5对组合均没有对‘兰州大接杏’和‘白木杏’这两个品种扩增出任何条带，表明要对中国杏品种资源的S基因型进行全面、准确鉴定，除了采用不同的引物组合外，还需要根据中国杏的S基因序列进一步开发、设计新的专用引物；③ EM-PC2consFD+EM PC3consRD和PruC2+PruC4R两对组合对‘红丰’、‘新世纪’、‘红荷包’和‘二花槽’等4个品种的扩增结果表明，依据本试验所提出的扩增系数和扩增成功率两个指标评价不同引物组合的扩增效果是有效的。     

Bestimmung der S-Allele in Brennkirschensorten (Prunus avium L.)

Zusammenfassung Brennkirschen sind Süßkirschensorten, welche spezielle Eigenschaften für die Branntweinproduktion und die industrielle Verarbeitung besitzen. Ihr Hauptanbaugebiet in Deutschland befindet sich in Baden-Württemberg und in Teilen Bayerns. In den letzten Jahren wurde über einen geringen Fruchtbehang in neu gepflanzten Brennkirschenanlagen berichtet. Süßkirschen sind selbstinkompatibel und benötigen deshalb eine Bestäubersorte. Ursache ist ein gametophytischer Selbstinkompatibilitätsmechanismus, bedingt durch einen S-Lokus. Mit Hilfe molekularer Marker wurden deshalb Untersuchungen zu den S-Allel-Kombinationen in 62 Brennkirschensorten bzw. -herkünften aus dem Raum Oberkirch, Mittelbaden, durchgeführt. Im Ergebnis konnten zwei bisher noch nicht beschriebene S-Allele und 14 neue Inkompatibilitätsgruppen ermittelt werden. Ursache für den schlechten Fruchtansatz in neuen Kirschenanlagen mit den Hauptsorten Dolleseppler und Benjaminer sind die gleichen S-Allel-Kombinationen (S₁S₇) in beiden Sorten. Als mögliche Befruchtersorten werden die Sorten Schwarze Schüttler (S₆S₇), Hartwälder (S₄S₇), Feuerkirsche (S₄S₁₄) und die ehemalige Sorte Dolleseppler der Baumschule Kiefer (S₁S₄) empfohlen.     

杏果实成熟期糖酸和色素物质含量的分析

以2个仁用杏和8个鲜食杏品种为试验材料,测定了果实成熟期果肉可溶性总糖、还原糖和可滴定酸,果肉和果皮花色苷、叶绿素、类胡萝卜素和类黄酮的含量。根据鲜食杏品种果肉的颜色,将其分为白色果肉组、黄色果肉组和橙黄色果肉组,比较了不同颜色果肉组间糖酸和色素物质含量的差异,并对果肉颜色与各指标间进行了相关性分析。结果表明:仁用杏品种果肉可溶性总糖含量低于鲜食杏,而可滴定酸含量高于鲜食杏;二者间还原糖含量无显著差异。花色苷仅分布于红色杏品种的果皮组织。除花色苷外,不同杏品种间,仁用杏品种果肉和果皮色素物质含量均高于鲜食杏品种;同一杏品种中,果皮色素物质含量均高于果肉。鲜食杏不同颜色果肉组间果肉可溶性总糖、可滴定酸和类胡萝卜素含量差异显著。果肉颜色与果肉类胡萝卜素含量极显著相关(r=0.872),与果肉叶绿素、类黄酮和糖酸含量则无显著相关性。     

Self- and cross-(in)compatibility between important apricot cultivars in northwest Iran

Summary

Knowledge of the self-(in)compatibility trait in commercial apricot cultivars is of great importance for breeders and growers. Five commercial apricot cultivars, widely grown in Iran, were self- and cross-pollinated to determine their pollen and stylar compatibility. Fruit-set in the orchard and pollen tube growth in pistils, from flowers pollinated in the laboratory, were evaluated. In addition, specific primers previously designed to amplify fragments of the S alleles responsible for the incompatibility trait, were used to amplify DNA extracted from the five cultivars.All results agreed and confirmed that three out of the five cultivars studied were self-incompatible, two of which were cross-incompatible and therefore had the same genotype. The cultivars, ‘Ghorban-e-Marageh’ and ‘Ghermez-e-Shahroodi’ were self-compatible and, interestingly, shared a PCR band with all Spanish self-compatible apricot cultivars examined to date.     

中国杏自交不亲和花粉特异SFB基因的鉴定与序列分析

利用花粉SFB基因的同源扩增结合内切酶酶切的方法,在9个中国杏品种中首次克隆了6个花粉SFB基因,对应花柱S-RNase基因序号将其分别命名为Par-SFB8、Par-SFB9、Par-SFB11、Par-SFB17、Par-SFB23和Par-SFB26,在GenBank上的登录号分别为EU652883、EU935588、EU652884、EU652885、EU652886和EU652887。序列分析表明杏的花粉SFB基因具有蔷薇科李属植物SFB基因的典型结构特征;其内含子位于5′端非翻译区,长度90 ~ 108 bp,核苷酸序列的同源性为63.9% ~ 93.3%。氨基酸序列的同源性比较表明,杏花粉SFB基因的种内同源性为73.7% ~ 85.3%;与李属其他植物花粉SFB基因的种间同源性为75.2% ~ 97.7%。利用特异PCR扩增进一步确定了杏的S9、S11、S17和S26单元型花柱S-RNase与花粉SFB基因间距离,表明花柱S-RNase与花粉SFB基因紧密连锁;同时组织特异性表达分析确定SFB基因在花粉组织中特异表达。以上结果说明获得的SFB基因为杏的花粉候选S基因。     

Somatic embryogenesis in cell suspension cultures of Prunus persica (L.)

Cell suspension cultures were derived from adventitious somatic embryos induced on cotyledons of rescued peach embryos. Embryogénie calli were induced and established on modified MS medium supplemented with 2,4-D, BAP, KN and inositol. Development of embryos during suspension cultures was identified by growth pattern and by histological and cytological examination, using acetocarmine stain. More embryogenic calli were produced using 2,4-D (5 (µM) than NAA. A gradual reduction of 2,4-D increased the recovery of globular embryos. Heart-stage embryos were developed on high-nitrogen salt medium, with added Ca(N03)2 (1000 mg I"1) and 6% sucrose. Somatic embryos turned green and sometimes red under white light. Embryo viability was highest when subculturing every 14 days. Viability of cells and differentiating embryos was assessed, microscopically, with Alexander’s stain. Viable embryos reacted pink to red while non- viable embryos reacted green to blue. Embryogenic cell suspension cultures were maintained for over ten months by regular subculturing after two weeks on modified MS medium with low 2,4-D, BAP and KN.     

不同碳源和氮源对杏斑点病病菌的影响

以杏斑点病病菌为试材,研究了不同碳源和氮源对杏斑点病病菌菌丝生长和孢子萌发的影响.结果表明:最适菌丝生长的碳源为乳糖、麦芽糖、蔗糖,其次为葡萄糖;最适宜菌丝生长的氮源为谷氨酸,其次为硝酸钙、硝酸钠、甘氨酸.最适宜孢子萌发的碳源为淀粉;最适宜孢子萌发的氮源为氯化铵.     

桃果实水孔蛋白cDNA的分离

使用逆转录聚合酶链失反应(RT-PCR)技术从桃果实的果皮组织中分离出来2个编码水孔蛋白部分cDNA克隆.比较结果表明,其推导的氨基酸序列与从其他一些植物体中分离出来的液泡膜内在蛋白有很高的同源性.数量PCR分析表明,编码水孔蛋白基因pWCl在果实发育早期表达,而pWC2在果实发育期间全过程均有表达.     

‘凯特’杏花粉离体培养条件筛选

以‘凯特’杏为试材,采用不同培养基组分及不同培养基类型的方法,测定了花粉萌发率和花粉管伸长长度,研究了不同培养基组分及不同培养基类型对花粉萌发和花粉管伸长的影响。结果表明:固体培养基适合‘凯特’杏离体萌发,最适培养基组分是10%蔗糖+0.10%硼酸+0.5%琼脂;在该培养基上,花粉萌发率达90%以上,花粉管生长长度为13 510.30μm。     

桃杂交后代主要经济性状遗传变异分析

对桃品种（系）１２个杂交组合１３６１个单系的后代主要经济性状的遗传变异情况进行了分析，包括Ｆ＿１开花期、果实成熟期、果实大小、颜色和风味、果形、果肉硬度、果肉颜色、丰产性、核粘离性、树体性状和综合品质等，为进一步合理配置亲本提供参考依据。     

Growth Promoting Substances in The Developing Fruit of Peach (Prunus Persica L.)

Growth substances extracted from the ovules of developing fruits of July Queen, Halehaven and Fall Late peaches–respectively early, midseason and late-maturing cultivars–were studied. Whole and fractionated extracts were bioassayed, and the growth of the fruit and the development of the seed were examined.

Determination of “total” growth-promoting substances during the development of the fruits resulted in concentration curves that were generally similar from year to year. The seeds of each cultivar, regardless of its season of maturation, produced a first flush of growth substances that seemed to be correlated with either nucellus or endosperm development. The second and major growth substance peak came at the time of rapid embryo growth and maximum endosperm development. At this time fruits of July Queen ripened and abscissed. A third flush of growth substance was detected at the same time in Halehaven and Fall Late seeds, at which time the embryo was essentially complete. The mature fruits of Halehaven then abscissed, but Fall Late fruits remained on the tree for another three or four weeks before maturation and abscission. It was concluded that the concentration of seed growth substances as measured here was not related to the final-flush of growth of peach fruits, or to the abscission of mature fruits.

In dissected ovules of Halehaven peaches the endosperm was the most concentrated source of total growth substances on a dry weight basis but, because of its small size, it contributed less of them on a per-seed basis than did the the integuments at first, and later the embryo. The nucellus was a minor source of growth substances, both on a concentration and on a per-ovule basis. Apparently, the total growth substances that constituted the second and major auxin peak in Halehaven peach ovules came mostly from the integuments, endosperm and embryo, whereas the third peak could be attributed almost entirely to substances from the embryo. These studies of individual ovule tissues were made after that period of development during which the first growth substance peak made its appearance.

Studies of growth substances in the flesh of developing peach fruits revealed only trace amounts with no consistent pattern, although there is some evidence that a true picture may not have been obtained because of interference by inhibitors.

Silica gel partition column chromatography showed the presence of several growth-promoting substances which changed qualitatively and quantitatively during the growing season. One of these was probably indole-3- acetic acid, its identification being based primarily on studies by gas chromatography and spectrophotofluorometry.     

甜樱桃品种吉列玛叶片再生不定梢的研究

在基本培养基NN69或WPM上附加BA和低浓度的NAA，IAA，IBA作培养基，诱导甜樱桃品种吉列玛试管苗的叶片产生了不定梢。以WPM附加BA 3mg/L和IAA0．3mg/L的处理，不定梢再生率最高为33．3％。     

果梅基因组DNA提取方法的比较及ISSR分析

以果梅品种鸳鸯梅、皇后梅和肖山选的幼嫩叶片为试材,提取果梅的DNA.针对果梅组织细胞体内含有较多的酚类、糖类及萜类等次生代谢物质的特点,采用传统的CTAB法、改良的CTAB法和改良的SDS法提取果梅基因组DNA,并比较其提取效果.结果表明:改良的CTAB法可获得高质量、高得率的基因组DNA,OD260/280比值在1.80左右.通过基因组DlNA-ISSR分析,完全满足试验要求.并进一步对改良的CTAB法的关键步骤作出具体的分析讨论.     

桃果肉颜色、果皮茸毛和花粉育性性状的分子标记

 以91-42-51 ×瑞光2号的杂交后代共48个单株为试材, 采用集群分离分析法(BSA) , 对桃果实的有毛/无毛、白肉/黄肉以及无花粉/有花粉3 个性状进行了分子标记研究。研究获得SSR 标记UDP96-018 (300 bp) 与有毛性状连锁, 遗传距离4.5 cM, SSR标记UDP98-407 (680 bp) 与白肉性状连锁, 遗传距离为2.2 cM; 根据拟南芥雄性不育序列标记设计的引物扩增出的两个片段NNJ-I (600 bp ) 和NNJ-I (900 bp) 与桃无花粉性状之间的遗传距离为0 cM。这些标记的获得为桃分子标记辅助育种提供了有效的筛选手段。