基于成熟期的桃品种遗传多样性SSR分析

以95份桃育成品种为试材,利用32对SSR分子标记对桃育成品种进行遗传多样性分析。结果表明:筛选的18对SSR引物共检测出93个等位基因,其中多态性等位基因为50个。桃群体的平均Nei’s基因多样度为0.5652,Shannon遗传表型指数为1.1125,说明桃总群体遗传变异偏低。针对桃的成熟期进行遗传分析,发现早熟和中熟类群遗传数据相似,推测二者亲缘关系相近;同理,推测晚熟品种与前二者关系较远。表明桃的成熟期与其等位基因相关,该研究结果可为进一步研究桃的遗传分析奠定基础。     

基于成熟期的杏品种遗传多样性SSR分析

以168份杏育成品种为试材,采用SSR分子标记方法,利用16对多态性好、稳定性强、带型清晰的特征引物,扩增等位基因,研究了不同杏树不同群体的遗传多样性,以期为杏种质资源鉴定和品种亲缘关系分析在分子水平上提供依据。结果表明:杏群体的Nei’s基因多样度平均值为0.715 8,Shannon遗传表型指数为1.497 3,说明杏的总体遗传变异偏高,品种多样性丰富。通过对杏的成熟期进行遗传分析,结果发现,早熟和中熟2个类群的遗传数据相近,推测可能存在相近的遗传关系,晚熟与早熟类群等位基因数据差异较大,说明二者亲缘关系较远。     

基于成熟期的梨品种遗传多样性SSR分析

以63个梨主栽品种为试材,采用SSR分子标记技术与毛细管电泳技术相结合的方法,研究了梨不同成熟期的种质资源遗传多样性,以期为梨栽培品种的品种鉴别、亲子关系鉴定、分类及系统关系研究、遗传图谱构建与标记辅助育种在分子水平上提供参考依据。结果表明:3份梨种质的观测等位基因数(Na)平均为14.8,有效等位基因数(Ne)平均为7.363 1,Shannon多样性指数(I)平均为2.195,期望杂合度(Nei)平均为0.845,说明梨总群体遗传变异偏高。依据梨系统将其分为白梨、秋子梨、日本梨、砂梨和西洋梨5个类群,其中白梨系统在Shannon多样性指数和期望杂合度上最高,分别为2.069 9和0.831 1,秋子梨系统最低,分别为0.420 8和0.303 6,白梨与砂梨类群的遗传一致度最高为0.715 2。     

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。     

黑龙江省黑木耳主栽品种SSR遗传多样性初探

从30对SSR引物中筛选出多态性较好的引物对黑龙江省黑木耳主栽品种进行SSR遗传多样性分析和主坐标分析,结果筛选出8对扩增多态性较好的引物,UPGMA法进行聚类分析,可将其分为三类,目前主栽黑木耳品种聚为一类,以前主栽黑木耳品种聚为一类,分离野生黑木耳聚为一类。供试黑木耳菌株遗传相似系数在0.60~0.97,表明遗传多样性较高,主坐标分析也可将其分为三类,这与UPGMA聚类结果一致。     

厚皮甜瓜遗传多样性的SSR分析

采用61对SSR特异引物对46份厚皮甜瓜材料进行分析,其中52对扩增出条带,47对引物具有多样性,扩增带分子量在150～1500bp,187条谱带中154条谱带具有多态性,多态率占82.35%,平均每个引物可扩增出3.60条带。同时,对材料网纹、果形、单果质量、有无条带、果皮颜色、果肉颜色、肉质、种子、有无麝香味、裂叶、是否自落和花性型等12个性状进行了田间调查,经过phylip软件和NTSYS软件分析后,计算出材料间遗传距离,并做出SSR分子标记和性状2个聚类图。结果显示,46份材料的遗传距离在0.0188～1.2119,而单一性状气味、果皮颜色、果肉颜色和表面网纹的遗传距离都小于0.1,十分接近;2种聚类分析结果存在较大差异,这与供试材料的亲缘关系十分复杂,大部分材料来源相近,具有相同的亲本,和回交亲本存在一定关系。     

3个生态群杏品种遗传多样性SSR分析

利用8对SSR引物对中亚生态群、华北生态群、欧洲生态群30个杏品种进行了分析。结果表明,在遗传相似系数为0.73时,供试材料分为3组,第1组包括欧洲生态群10个品种和中亚生态群2个品种;第2组包括华北生态群10个品种和中亚生态群5个品种;第3组包括中亚生态群3个品种。其中3对引物(aprigms1、aprigms6、UDP96-018b)在中亚生态群杏中扩增出了特殊基因,1对引物(UDP96-003b)在华北生态群杏中扩增出了特殊基因。中亚生态群杏品种的遗传多样性最为丰富,与华北生态群杏品种亲缘关系较近,可能是我国栽培杏的起源中心。     

基于SSR标记的现代月季遗传多样性研究

以来自亚洲、欧洲、北美洲的51个现代月季品种为试材,采用18对多态性高的SSR引物进行分析,研究不同起源地的现代月季品种的遗传多样性、遗传结构及遗传距离。结果表明:共检测出64个位点,多态位点比例为96.8%;其中来自欧洲地区的多态位点比例最高,达到82.8%,遗传变异由高到低次序是欧洲月季、北美洲月季、亚洲月季;类群内的遗传分化(91.9%)大于类群间的遗传分化(8.1%),说明类群内变异是其主要变异来源;欧洲地区与北美洲地区现代月季的遗传关系较近。     

甜瓜种质资源遗传多样性的SSR 分析

利用SSR 分子标记技术对72 份甜瓜种质资源及育种材料进行DNA 指纹分类及亲缘关系研
究,20 对SSR 引物共扩增产生76 个多态性位点,平均每对引物产生3.8 个多态性位点。聚类结果将72 份
甜瓜种质资源分为7 个类群,其中类群I 包括新疆本地资源及几乎全部自育甜瓜种质资源64 份材料,这
说明本试验所选用的新疆厚皮甜瓜种质资源遗传背景比较狭窄。     

蟠桃种质SSR标记的遗传多样性分析

以38个蟠桃品种和9个其他类型桃品种为试材,利用24对位于桃参考图谱上8个连锁群的SSR引物进行了蟠桃种质资源遗传多样性研究。24对SSR引物共获得179个扩增位点,其中多态性位点171个,多态率达95.53%。蟠桃资源平均Nei’s基因多样性指数(He)为0.242 5,平均Shannon信息指数(I)为0.379 8;南方蟠桃品种群多态性位点百分率为74.30%,Nei’s遗传多样性指数为0.203 8,Shannon信息指数为0.316 3;北方蟠桃品种群的遗传多样性相对较高,多态性位点百分率为91.06%,Nei’s遗传多样性为0.244 9,Shannon信息指数为0.382 4。群体间存在较小的基因分化系数(Gst=0.065 9),遗传变异有很大一部分是来自群体内,可能与其较大的基因流Nm=7.086 1有关。UPGMA聚类分析结果表明,相似系数为0.66～0.95,在相似系数为0.67时,所有南方品种聚于同一类群,北方品种除新疆蟠桃、黄肉蟠桃及香金蟠外也聚于同一类群,聚类结果支持蟠桃多起源假说。     

桃幼胚离体培养再生植株的研究

 以中晚熟桃品种‘京艳’、‘绿化9 号’、‘ 晚蜜’和‘大久保’为试材, 初步建立了桃体细胞胚发生和增殖的三阶段程序, 即胚性愈伤组织诱导、体细胞胚发生和发育、体胚萌发成株以及次级体细胞胚的增殖。各品种体细胞胚的萌发成株率分别为73. 5 %、38. 6 %、5. 4 %和58. 2 %。用苯脲类细胞分裂素TDZ有效诱导‘京艳’的幼胚子叶直接再生植株, 再生率70 % , 平均每外植体不定芽数13. 8 个, 再生不定芽生根后获得完整植株。     

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。     

Genetic diversity in Apium graveolens and related species revealed by SRAP and SSR markers

Genetic diversity is essential to crop improvement. However, lack of molecular markers prevents the understanding of genetic diversity in celery and related species. In this study, SRAP and SSR markers was firstly used to evaluate genetic variation in 68 accessions of Apium graveolens and related species. A total of 888 bands were generated by 40 primer combinations of SRAP and 32 bands were produced by eight SSR markers. Of the 920 bands, 95.1% were polymorphic between A. graveolens and related species, and 49.2% were polymorphic within A. graveolens. Cluster and structure analysis could distinguish local celery, celery, and related species. These results suggested that both SRAP and SSR technologies can be efficiently used to characterize genetic variation and analyze genetic relationship in celery and related species.     

桃品种耐寒性研究

用电导法、萌芽生长法和组织变褐法对82个桃品种或优系的耐寒性评价,结果表明：桃一年生休眠枝的耐寒温度为-19．0～-27．0℃,叶芽为-18．5～-26．1℃,花芽为-17．0～-24．0℃;枝、叶芽和花芽三者间耐寒性呈显著正相关,抗性强弱的顺序为：枝＞叶芽＞花芽;枝组织的耐寒力顺序为：韧皮部＞木质部＞形成层＞髓;品种间、品种群间及变种间抗寒性也有不同程度的差异。提出在很少发生-23．0℃以下低温的地区为桃适宜经济栽培区。     

板栗野生居群遗传多样性研究

使用从板栗中提取的10个微卫星引物对4个中国栗野生居群的69个个体进行扩增,共检测到84个等位基因,每个位点的等位基因数为4～13,每位点的平均等位基因数目为8.4个,平均有效等位基因数(Ne)为4.998,平均期望杂合度(He)为0.777,平均PIC值为0.739,Nei’s多样性指数(h)为0.771,以陕西汉中居群具有最高的遗传多样性。遗传分化系数Gst仅为0.141。UPGMA树状聚类图表明4个野生群体被分为两部分,其中云南、四川、安徽3个居群距离相近位于同一组,此结果与PCA分析结果一致,2种方法均显示居群间的遗传关系与实际地理分布不完全相关。     

桃幼胚下胚轴高频植株离体再生

以桃品种京艳和晚蜜不同发育阶段合子胚下胚轴为外植体进行离体培养,用6-BA、CPPU和TDZ等细胞分裂素诱导下胚轴直接再生不定芽试验。结果表明,花后70d下胚轴为诱导的最佳材料,供试细胞分裂素中,诱导活性为TDZ>CPPU>6-BA,苯脲类细胞分裂素TDZ5mg/L与NAA0.01mg/L配合对下胚轴再生具有极强的诱导作用,京艳诱导再生率高达95.2%,平均不定芽数达每外植体25.3个。京艳的诱导率显著高于晚蜜。IBA 0.5mg/L适于诱导再生幼茎的生根,生根率和移栽成活率分别达到92.6%和65.5%。组织学观察表明不定芽起源于皮层薄壁细胞,属直接再生方式。     

S_allele identification and genetic diversity analysis of apricot cultivars

In this study, the sexual incompatibility and S-allele diversity of 24 Turkish apricot cultivars, Paviot and Sak?t-1 as parents and 127 F₁ progenies were identified by polymerase chain reaction (PCR) and sequencing techniques. Additionally, genetic diversity and relatedness among the 24 cultivars were determined using 18 simple sequence repeat (SSR) markers from the genus Prunus. PCR for S_alleles identified nine different S-RNase alleles in the 24 apricot cultivars, namely S_c, S₁, S₂, S₈, S₉, S₂₀, S₂₄, S₅₂, and S₅₃. All primers amplified only one S_allele in the cultivars Adilcevaz-1, Adilcevaz-3, Ethembey, Pasamismisi, Canakkale, and Soganci. Most of the Turkish cultivars were self-incompatible. The S_c allele was present in only three cultivars (Canakkale, Ethembey, Imrahor) that are, therefore, self-compatible. The S_alleles of cultivars Paviot and Sak?t-1 displayed homology with the S_c, S₂ and S₂₀, and S₅₂ alleles. In the 127 F₁ genotypes, the two S_alleles of Paviot were inherited by roughly half of the offspring, while about 76% of the offspring inherited the S₅₂ allele from Sak?t-1, and less than 24% inherited S₂₀. The amplification using all SSR 18 primers was successful and produced 128 polymorphic alleles with an average of 7.11 alleles per locus. Among the apricot cultivars studied, expected heterozygosity (He) ranged from 0.33 to 0.72, observed heterozygosity (Ho) ranged from 0.42 to 1.00, PIC values were between 0.28 and 0.89, and similarity rates were between 0.30 and 0.68. The cultivars Levent and Ozal were genetically closest (0.68) while cultivars Sak?t-3 and Soganc? were the most distinct (0.30).     

Isolation,cloning, and expression of five genes related to nitrogen metabolism in peach (Prunus persica L. Batsch)

An optimal dosage of nitrogen has long been known to play important roles in governing nitrogen-use efficiency and improving the yield of plants. However, there is limited information on the isolation and expression profiles of genes related to nitrogen metabolism in peach (Prunus persica L. Batsch). This study isolated five genes involved in nitrogen metabolism and evaluated the effects of a single foliar application of urea on levels of expression of these genes in the leaves of ‘Dazhenbaochiyue’ peach. Cloning resulted in the isolation of 1,767, 1,236, 1,074, 1,758, and 2,721 nt-long cDNAs with full-length open reading frames encoding 588, 411, 357, 585, and 906 amino acids representing the asparagine synthetase (AS), glutamate dehydrogenase (GDH), glutamine synthetase (GS), nitrite reductase (NiR), and nitrate reductase (NR) genes in peach, respectively. An alignment of multiple amino acid sequences revealed that the AS, GDH, GS, and NR proteins in peach shared high levels of sequence conservation or identity at the amino acid level with their homologues in Arabidopsis thaliana and grapevine (Vitis vinifera). Based on analyses of expression of the five genes in peach leaves, we demonstrated that genes related to nitrogen metabolism responded to a foliar application of urea within 2 d. This was consistent with previous reports which indicated that leaves rapidly absorbed urea-nitrogen after foliar application. Foliar application of 0.5% (w/v) urea inhibited expression of GS and NiR, but increased expression of GDH, AS, and NR compared to control leaves. The different patterns of expression of the five genes in this study suggested that their expression might reflect their various roles in nitrogen metabolism, plant N status, and responses to weather conditions such as high light intensity, temperature, or humidity, all of which affect photosynthesis. These findings provide a basis for future functional analyses of these five genes in peach.     

中国樱桃14个自然居群遗传多样性和遗传结构的SSR评价

以中国樱桃14个自然居群280个个体为材料,采用SSR分子标记技术分析其遗传多样性和遗传结构。结果显示：11对SSR引物共检测到80个等位基因,各引物扩增条带在4 ~ 13条之间。基因多样性指数（h）为0.5431 ~ 0.7151,Shannon’s信息指数（I）为0.9057 ~ 1.4684。分子方差分析（AMOVA）表明,遗传变异主要来自居群内（70.00%）,Mantel检验显示总群体的遗传距离和地理距离显著相关（r = 0.472,P = 0.011)。因此,中国樱桃在居群水平（PPL = 100%,h = 0.643,I = 1.207）和物种水平（PPL = 100%,h = 0.743,I = 1.591）上均具有较高的遗传多样性。     

Evaluation of the genetic diversity of Asian peach accessions using a selected set of SSR markers

A total of 94 peach accessions from the Zhejiang province of China were analyzed using 34 polymorphic single-locus simple sequence repeat (SSR) markers. Genetic distance analysis divided the accessions into two major clusters, one with mainly local accessions from the Fenghua region. Preselected lines from some crosses were exclusively clustered with the local ones, possibly related to maintaining the taste quality of the fruit. The number of alleles per locus at most loci was two or three, with an average of 2.85. The value of observed heterozygosity varied from 0.05 to 0.84 (average of 0.48). Diversity within the introduced accessions was higher than that of the Fenghua local accessions. Of the accessions analyzed in this study, 94% were individually identified. Those that could not be differentiated were all derived from the ‘Yulu’ cultivar, being either mutations or of identical origin.