番茄苗期耐盐相关遗传位点鉴定及分子标记开发

以501份番茄重测序自然群体为研究材料,于苗期测定Na⁺和K⁺含量并计算Na⁺/K⁺比值,通过全基因组关联分析及单倍型分析,筛选耐盐相关显著关联位点,鉴定基于显著关联位点不同基因型材料的耐盐表型,利用该位点开发分子标记及验证。结果表明：大果番茄的驯化和改良过程导致了植株地上部Na⁺含量和Na⁺/K⁺比值的降低。通过全基因组关联分析在7号染色体上鉴定到与番茄地上部Na⁺含量和Na⁺/K⁺比值显著关联位点NTC7(Na⁺tolerance loci on chromosome 7),该位点包含两个功能已知的耐盐基因,HKT1;1和HKT1;2。序列分析和单倍型分析鉴定到HKT1;2启动子上存在1个6bp插入缺失与地上部中Na⁺含量和Na⁺/K⁺比值高度关联,通过盐处理验证该位点与番茄耐盐...     

西瓜果实形状的分子精准鉴定

为了实现西瓜果实形状的分子精准鉴定,利用"国家西瓜甜瓜中期库"的资源和高通量测序平台,利用两个不同的杂交分离群体(F_2和BC_1P_1)分别鉴定到1个159 bp插入缺失和1个非同义SNP导致的果形突变(由圆果形变为长果形),且SNP突变在159 bp插入缺失的基因组区域内。利用这个SNP开发CAPS标记Markersun在两个群体和128份西瓜种质中进行分析,发现标记Markersun在两个群体中与果形表型共分离;在西瓜种质中可同时区分插入缺失和SNP的变异,且与果形表型共分离。同时这两个变异与果形的共分离在196份西瓜核心种质的基因型分型中也得到验证。此外,通过对不同类型西瓜种质的基因型分析发现,SNP变异导致的长果形出现的时间更早且独立遗传,而159 bp插入缺失引起的长果形是栽培西瓜驯化过程中产生的。本研究中首次利用不同类型的西瓜核心种质实现了西瓜果形的分子精准鉴定,还挖掘到两个西瓜果形的功能变异并开发标记,为西瓜果形的分子精准鉴定提供技术支撑,同时为果形性状基因功能验证提供靶标,加速了西瓜果形性状基因的调控机理研究。     

基于SLAF-seq技术的不同类型越橘遗传变异和群体结构研究

【目的】利用越橘属栽培品种及野生资源在全基因组范围内筛选差异SNP分子标记，分析其遗传变异，了解不同类型群体间的遗传变异、基因交流、群体结构，为越橘种质创新提供遗传背景依据。【方法】以越橘属9个种的62份种质资源为材料，利用特异性位点扩增片段测序技术(SLAF-seq)获得的基因组SNP标记比对到品种Draper参考基因组上，进行供试材料遗传研究。【结果】测序获得169.87 Mb reads数据，平均Q₃₀值94.99%，包含172 513个多态性SLAF标签，筛选到高质量SNP标记4 133 595个，均匀分布在越橘12条染色体上。基于SNP标记的系统进化树准确反映了供试野生、北高丛和南高丛类群遗传差异以及品种间系谱遗传关系，反映出野生类型与栽培品种遗传差异显著，供试野生种具有独立的遗传背景且种间无基因交流发生，但野生种到栽培种存在单向的基因流；栽培品种类群存在明显的亚群结构，亚群间存在普遍的基因交流；半高丛品种北陆和南高丛品种军号与北高丛类型紧密聚类，矮丛品种美登、半高丛品种北村、黑珍珠遗传背景倾向于野生种。【结论】栽培品种近亲杂交和骨干亲本高频率使用导致品...     

苦瓜植株卷须发生的初始节位及其分叉的遗传分析

以栽培苦瓜自交系佛沥112 和野生苦瓜自交系THMC170 为亲本,构建4 个世代（P₁、P₂、F₁、F₂）遗传群体,采 用主基因+ 多基因混合遗传模型对苦瓜植株卷须发生的初始节位进行遗传分析;以栽培苦瓜自交系佛沥734 和半野生苦瓜自 交系AVBG1602 为亲本,构建4 个世代（P₁、P₂、F₁、F₂）遗传群体,对苦瓜植株卷须的分叉性进行遗传分析。结果表明, 苦瓜植株卷须发生的初始节位遗传符合2 对等显性主基因+ 加性- 显性多基因遗传模型（E-6 模型）,其中主基因遗传率为 15.81%,多基因遗传率为7.83%。苦瓜植株卷须的分叉对不分叉受1 对显性基因控制,其中卷须分叉表现为显性,卷须不分 叉表现为隐性。研究结果为深入解析苦瓜植株卷须发生及发育的遗传机制奠定了基础。     

45份宽皮柑桔野生和地方资源遗传多样性分析

利用SSR、InDel及来自于lncRNA的分子标记对不同地区的45份野生、半野生及地方宽皮柑桔资源进行了遗传多样性分析。31对引物共获得81个多态性标记位点,多态性标记位点的平均香农多样性指数为0.725,平均期望杂合度为0.413。群体遗传结构、主坐标轴和聚类分析均表明,莽山野柑是一类原始、独特的柑桔种质;起源于南岭山脉的野生宽皮柑桔,如悬崖野桔、道县野桔、江永野桔-1、莽山桔、崇义野桔和江永野桔-5等构成了一类相对独特的原始野生类群;古老栽培柑桔中的皱皮柑类型,包括旺苍皱皮柑、皱皮柑、藏桔等构成了另一个原始古老类群,现代栽培柑桔城固冰糖桔中具有典型皱皮柑基因的渗入;宽皮柑桔中的一些育成品种及杂柑资源,具有较丰富的遗传多样性。本研究证实了起源于南岭山脉一带的野生宽皮柑桔在遗传背景上较为接近,其多样性也低于现代栽培宽皮柑桔,缺乏人为驯化和外源基因渗入,为南岭山脉是我国宽皮柑桔的起源中心这一论断提供了科学依据。由于长期的人为驯化和外源基因的渗入,宽皮柑桔的演化伴随着遗传多样性水平由低向高逐渐演变的过程。     

43份云南野生枇杷种质遗传多样性与亲缘关系分析

应用SSR、SRAP等2种分子标记方法,分析了43份云南野生枇杷种质的遗传多样性及其与部分栽培品种、野生近缘种的亲缘关系。结果表明：云南野生枇杷种质具有较丰富的遗传多样性,其多态性位点百分数为91.32%,平均期望杂合度为0.3165,平均香农多样性指数为0.4753;在相似系数0.751水平上,43份材料可被划分入4个大类,其中类Ⅲ在相似系数0.816处可细分为6个亚类;主成分分析则更为直观地展现了种质间的亲缘关系。聚类结果与种质类型相关,并呈现出一定的地域性分布规律。结合形态学特征,鉴定该批云南野生种质以栎叶枇杷种内类型居多,甚至有出现变种的可能。     

青花菜相同亲本的DH 与F2 群体遗传多样性的比较

 以表型差异明显的两个青花菜（Brassica oleracea var. italica）高代自交系为材料杂交获得F₁，通过游离小孢子培养技术和人工自交方法，分别构建了由176个DH系组成的DH群体和由176个单株组成的F₂群体。采用性状分离分析和群体遗传多样性分析方法，对两个群体13个主要农艺性状的平均值、变异系数及性状分离区间等遗传表现进行分析。结果表明：13个主要农艺性状均是受多基因控制的数量性状，在DH群体和F₂群体中均发现了正向和负向两个方向的超亲基因型，两群体的平均变异系数和遗传多样性指数相近。可见，通过DH 技术育种不仅可以加快亲本的纯化进程，而且可获得广泛分离的育种资源，从而提高育种效率。     

一个控制现代栽培番茄果形、果实大小和市场性状的新基因

正番茄果实的形状和大小是极为重要的性状。在野生番茄人工驯化的过程中,目前已知控制果实形态的主效基因包括:SUN、OVATE、LC、FAS、FW2.2、FW3.2和FW11.3等。然而人们对栽培品种果实形状和大小变异的遗传学基础知之甚少。之前的一项研究对市场上不同果实形态的大果番茄自交系的6个性状位点(sun、lc、ovate、fas、fw2.2、fw3.2)进行了评估,并没有发现任何变异,     

大白菜BrAOP2 基因的遗传多样性及其与硫甙含量的相关性分析

以大白菜品种Chiifu 为试验材料, 采用RT-PCR 技术, 克隆了3 个硫甙合成关键基因
BrAOP2 基因的cDNA 序列（BrAOP2.1、BrAOP2.2、BrAOP2.3）。通过氨基酸同源性比对分析,BrAOP2.1
与BrAOP2.2 的同源性为78%,BrAOP2.1 与BrAOP2.3 的同源性为74%,BrAOP2.2 与BrAOP2.3 的同源
性为81%。通过32 份大白菜重测序数据分析了3 个BrAOP2 基因CDS 序列的遗传多样性,并对其与硫甙
含量的相关性进行了分析。结果表明：在32 份大白菜中共检测到7 种硫甙,其中4-戊烯基硫甙（GBN）
含量和2-羟基-3-丁烯基硫甙（PRO）含量较高,分别占总硫甙含量的23.99% 和23.16%。在BrAOP2.1
基因编码区检测到5 个变异位点,在BrAOP2.2 基因编码区检测到7 个变异位点,在BrAOP2.3 基因编码
区仅检测到1 个变异位点。其中BrAOP2.1 基因的A1051G 变异位点与PRO 含量极显著相关,BrAOP2.2
基因的A560G、C753T、A790G 和T927C 变异位点与PRO 含量显著相关,BrAOP2.2 基因的G825A 位点
与4-羟基-3-吲哚基甲基硫甙（4OH）含量显著相关。     

甘蓝无蜡粉亮绿突变体材料LD10遗传规律及分子标记研究

为了明确结球甘蓝无蜡粉亮绿突变体材料LD10 的特征特性及遗传规律,测定了LD10 的主要农艺性状和营养物质
含量;对LD10 和普通材料21-3 的叶面超微结构进行了观察比较;以LD10 和21-3 为亲本构建6 世代群体,对LD10 无蜡
粉亮绿性状遗传规律进行分析。结果表明：无蜡粉亮绿突变体材料LD10 叶面蜡粉呈小颗粒状结构,缺失严重,而普通材料
21-3 叶面蜡粉呈长棒状结构,含量丰富,二者具有明显差异;LD10 的无蜡粉亮绿性状由1 个隐性基因（cgl-4）控制,其与
SSR 标记LT-SSR16 紧密连锁,遗传距离为4.7 cM。用LD10 与另一单基因隐性遗传的无蜡粉亮绿突变体材料10Q-961 杂交
获得F1 群体,F1 群体单株全部表现为叶面有蜡粉,推断LD10 与10Q-961 的无蜡粉亮绿基因位于不同的遗传位点。     

Use of chloroplast microsatellite markers as a tool to elucidate polymorphism,classification and origin of Tunisian grapevines

Chloroplast microsatellite markers were used in this study to genotype 43 grapevines accessions grown in Tunisia. Size variation was observed for the three cpSSR loci, both in the sample of cultivars and in wild accessions. The seven alleles observed in the sample of cultivars for the three loci are present in wild accessions except that their distribution is different. Levels of genetic diversity obtained for the Tunisian grapevines either in wild or cultivated gene pools are high and comparable with values obtained with other studied samples of Vitis vinifera. The distribution of haplotypes within the two samples is differential. Indeed, the chlorotype A is most abundant in the wild sample, whereas the chlorotype C is majority in the sample of cultivars. Haplotypes frequencies for cultivated grapevine distinguish haplotypes B and C as the most frequent (28% and 44% respectively) and haplotypes A and D as the least frequent (16% and 12% respectively). For wild grapevines, the seven alleles combined in three haplotypes, A, C and D. The haplotype A is the most frequent (44%) in the analyzed sample of wild accessions while haplotypes C and D show a frequency of 28%. Chlorotype distribution in Tunisian cultivars is comparable with that of cultivars in the Eastern Region representing the primary centre of domestication of the species. These results agree with the higher relevance of table grape cultivars in Tunisian viticulture and support an oriental origin of a large part of autochthons cultivars. Our results agree with other studies based in nuclear and chloroplast microsatellite markers and suggest independent domestication events for V. vinifera L. species.     

枇杷属植物落黄叶片颜色遗传多样性分析

调查发现枇杷属(Eriobotrya)植物种质落黄叶片有红褐色、金黄色、紫红色等24种代表性色泽。通过便携式色差仪观测枇杷属10个种176份种质落黄叶片正面和背面的色泽参数,探讨落黄叶片颜色的遗传多样性特征。结果表明:(1)枇杷属植物落黄叶片正面和背面的色泽参数L、a、b值变异系数差异较大,变幅为8.65%~185.30%,Shannon’s遗传多样性指数为1.95,说明落黄叶片色泽变异多样;(2)不同种质类群间和不同来源地类群间的变异系数和Shannon’s遗传多样性指数差异均较明显,呈现出栽培驯化程度越高,落黄叶片正面和背面的b值变异系数越小和a值变异系数越大的趋势;云南、贵州、福建地区的类群多样性指数较高;(3)系统聚类将落黄叶片正面色泽划分成12组,背面分成10组,且各组内所包含的枇杷种质类型差异大;(4)L值、b值对落黄叶片正面和背面的系统聚类分析贡献率均较大,落黄叶片正面L值与背面b值呈极显著正相关;(5)落黄叶片颜色的遗传多样性可作为枇杷属植物形态学特征描述的补充,为枇杷属植物的鉴别和利用提供新的参考,对枇杷分类研究具有潜在价值。     

从DNA水平上分析香菇不同菌株的遗传差异

本文选取了10个中国香菇野生种和2个栽培种.采用RAPD技术进行遗传差异测定.16个随机引物共扩增出118条DNA带,在此基础上分析并构建了遗传相关聚类图.结果表明,我国的香菇野生资源存在着较大的遗传差异,10个野生种菌株之间的遗传相似性在50%～80 %之间.     

Genetic diversity and relationships of wild and cultivated olives at regional level in Spain

Genetic diversity and relationships between local cultivars and wild olive trees from three important Spanish olive-growing regions, Andalusia (South), Catalonia and Valencia (from Eastern Mediterranean Coastal area), were studied by means of eight SSR loci. Distinct allelic composition and heterozygosity levels were found in wild olive populations and cultivars. The observed patterns of genetic variation revealed: a) the independent clustering of Andalusian wild olives in a separate gene pool, b) the belonging of wild populations and most cultivars from Catalonia to another gene pool, c) the joined clustering of Andalusian and a set of Valencian cultivars in a third gene pool, and d) clustering of wild individuals from Valencia to the three different gene pools. These results suggest that wild populations of Andalusia may represent true oleasters, the ones from Catalonia may be feral forms derived from cultivar seed spreading, while the population of Valencia seems to be the most admixed one. The significant differentiation between Andalusian and most Catalonian cultivars is indicating an independent selection of olive cultivars in the two regions. The detection of a certain wild genetic background in some Catalonian and Valencian cultivars and the similarity found between wild and cultivated forms may suggest the use of local wild trees in olive domestication. The proposed scenario for the development of olive cultivars in Andalusia includes an empirical selection of outstanding local wild genotypes followed by various generations of crosses and various replanting campaigns, as well as possible introductions of ancestral cultivars. Therefore, our findings would lead us to support the hypothesis that the current diversity found in Spanish olive cultivars may be regionally differentiated and due to both, autochthonous and allochthonous origin. The information obtained in this work gives insights into the genetic resources of the main olive producing country, demonstrating that wild olive populations and local cultivars both represent potential sources of useful variability for olive breeding programs.     

牡丹组野生种间亲缘关系和栽培牡丹起源研究进展

系统总结了近年来有关野生牡丹种间亲缘关系的研究进展,详细回顾了有关栽培牡丹起源的时间、地点、方式及祖先种等方面研究所取得的现有证据,并分析和讨论了栽培牡丹“多地”、“多元”起源的观点,认为栽培牡丹的起源途径是最初通过野生种的直接驯化获得原始品种,在此基础上通过突变和人工选择发展出古代和传统品种。除衍生出古代和传统品种的野生祖先种外,其它野生种通过自然与(或)人工杂交的方式参与现有品种群(亚群)的形成和发展。为寻找栽培牡丹起源的有力证据,不同栽培品种群(亚群)与野生种的分子系统学关系是今后研究的重点。     

贵州野生枇杷资源遗传多样性的ISSR分析

采用ISSR标记,构建了贵州39份野生枇杷种质的DNA指纹图谱,并对其遗传多样性进行评价。结果表明,以筛选出的15条引物进行PCR扩增,获得清晰、重复性好的谱带125条,其中多态性谱带108条,多态性比例为86.4%;应用引物815、825和841的组合,构建了39份枇杷种质的ISSR指纹图谱,可以有效区分所有供试材料;15条引物的谱带数据聚类分析表明,供试种质的遗传相似性系数为0.40～0.98,在相似性系数为0.70时,可将供试种质分成五大类;聚类结果与种质表型相关,与地理分布的相关性不明显。     

Screening wild Eriobotrya species as rootstocks for loquat

Summary

Six wild species of Eriobotrya were evaluated as rootstocks for domesticated loquat. Twelve-month-old seedlings of Oak leaf loquat (an Oak Leaf loquat ‘Zaozhong No. 6’ hybrid) and wild Fragrant loquat (E. fragrans Champ) had higher root-to-shoot fresh weight (FW) ratios than other wild species. When the domesticated loquat, ‘Zaozhong No. 6’, was grafted onto seedlings of six different wild loquat species, differences in compatibility were observed. Graft compatibility was lower for all combinations compared to ‘Zaozhong No. 6’ grafted onto domesticated loquat seedlings. When wild Henry loquat was used as a rootstock, delayed incompatibility was observed. Differences in soluble sugar contents (SSC) between scion and rootstock were observed, and there was a significant negative correlation between graft compatibility and the difference in SSC values. The influence of the rootstock on anatomical changes at the graft union were observed using a scanning electron microscope (SEM). Wild rootstocks also influenced scion growth, fruit bearing, and fruit quality. Graft combinations which used wild Fragrant loquat as the rootstock had increased cold tolerance and improved scion growth. Wild Fragrant loquat therefore appears to have value as a rootstock for domesticated loquat.     

Germplasm diversity and genetic relationships among walnut (Juglans regia L.) cultivars and Greek local selections revealed by Inter-Simple Sequence Repeat (ISSR) markers

Inter-Simple Sequence Repeat (ISSR) markers were used for the assessment of genetic diversity among walnut (Juglans regia L.) selections from Greek native populations in comparison to internationally cultivated walnut genotypes. Similarity coefficient values from 0.13 to 0.93 (with an average of 0.48) were found among the 56 accessions examined, which indicated the presence of a high degree of genetic variability. Most international cultivars were grouped together while most Greek native populations could not be placed into a distinct group. The Greek native population genotypes were found more diverse than the international cultivars. The mean similarity coefficient values for the former and latter were 0.44 and 0.56, respectively. In the cultivar group, two subgroups were distinguished; one consisted of genotypes involving ‘Payne’ and the other ‘Franquette’ in their pedigrees. Some cultivars and populations could not be grouped according to their pedigrees or collection area. Analysis of molecular variance (AMOVA) revealed that a larger part of the genetic variation exists among Greek walnut populations within a collection region (89%) than among the regions (11%). The pairwise regional PhiPT values indicated that the most geographically distant regions are the most genetically differentiated. The high variability existing in the Greek germplasm in combination with their valuable agro-morphological traits suggested that it would be beneficial to utilize this native germplasm pool in walnut breeding programs and germplasm management activities to maximize genetic diversity in cultivated walnut.