功能标记及在品种鉴定和辅助育种中的应用前景

功能标记是根据功能基因内部引起表型性状变异的多态性基序开发出来的一种新型分子标记.由于来自基因内的功能性基序,此类标记不需要进一步验证就可以在不同的遗传背景下确定目标等位基因的有无.本文详细描述了功能标记的概念及与其它几种类型遗传标记相比存在的优势:提出了功能标记开发的基本条件和发展策略.利用关联分析方法和近等基因系途径可以获得直接或间接的功能标记.结合本单位的研究方向,探讨了功能标记在分子标记辅助育种和品种鉴定中的应用潜力和开展相关工作的一些设想.     

我国北方地区花生品种的遗传多样性分析

利用SSR分子标记对我国北方地区育种品种以及区试品种共计68个栽培花生品种进行遗传多样性分析,结果表明,104对SSR引物中有15对在品种间存在多态,多态率为14.4%,平均每个标记产生3.8个位点。15对引物的多态信息量在0.397~0.667之间。山东地区花生品种的遗传多样性要高于我国北方其他地区供试品种。小粒组品种的遗传多样性要略高于大粒组品种。聚类分析表明我国北方68个育成和区试品种的遗传相似系数在0.64~0.91之间,分成两个大类,第一个大类可分成四小类,其中第三小类又可以分成六个亚类。这些结果表明我国北方地区现有育成和区试品种的遗传多样性较低,为拓宽花生育种的遗传基础,应从中发掘出有利用价值的品种。     

分子标记及其在核桃种质资源研究中的应用

介绍了分子标记技术的类型,以及目前常用的几种分子标记技术(RFLP、AFLP、RAPD、SSR、ISSR等)的基本原理、特点,综述了分子标记技术在核桃种质资源研究中的应用情况,并对目前分子标记研究存在的问题及前景作了分析。     

原位和异位保护对水稻种质资源遗传多样性的影响*

 利用24对SSR引物对云南省保山市分别在原位和异位保存条件下的12个水稻品种进行遗传多样性分析。24对引物在原位居群共检测到等位基因（A）110个,平均等位基因数(Na)为4.58个,平均有效等位基因数(Ne)为3.08个,其中多态性位点百分率(P)为91.76%,香农指数(I)为1.17;异位居群共检测到等位基因88个,Na为3.67个,Ne为2.52个,P为100%,I为1.03。结果显示,原位居群的各遗传多样性参数均高于异位保存居群,这表明与异位保护相比,原位保护是维持种群遗传多样性的一种更好的方式。     

云南屏边大围山微型鸡遗传多样性*

 云南屏边大围山微型鸡是我国具有独特遗传特性的、属于亚热带热带型早熟地方家禽品种。研究采用了33个家鸡特异性的微卫星标记对该鸡种自然群体中30个个体进行了多态性电泳检测,目的在于阐明其群体遗传变异和遗传结构状况。研究结果显示：33个微卫星座位共检测到65个等位基因,每个座位的等位基因数在1～3个之间,这些等位基因的频率大小在0.0333～1.000之间,平均每个座位的等位基因数为1.9697个,有效等位基因数在100～263个之间, 平均有效等位基因数为1.6846。群体平均杂合度（H）和群体平均多态信息含量（PIC）为0.1737和0.3279。结果表明,大围山微型鸡群体平均杂合度和群体平均多态信息含量较低,纯合度较高,未受到其它外来血液的混杂,是一个封闭的原始群体,具有较高的遗传一致性     

云南地方香稻与非香稻遗传多样性比较

1材料与方法 1．1材料 供试水稻种子全部来自于云南省农业科学院种质资源保存库,是否为香稻品种均依据保存目录的性状记载、并采用咀嚼法对其进行再次验证。其中香稻10份,非香稻45份（表见第57页Table 1：品种及其来源）。供试所有香稻和非香稻品种均具有不同的云南省种质资源保存库编号。特别是10份香稻品种,来自于云南的5个相距较远的地方,在云南栽培香稻中具有一定代表性。     

转EGFP基因猪胎儿神经干细胞的体外分化

 【目的】获得转EGFP基因神经干细胞并监测其自我更新、增殖和多向分化潜能;通过用EGFP对神经干细胞进行标记和体外追踪实验,为EGFP作为示踪标记对神经干细胞进行体内移植研究奠定基础。【方法】利用神经干细胞培养技术体系,从胎龄30 d猪胎儿脑组织中分离培养神经干细胞,通过脂质体介导转染技术,将EGFP基因导入神经干细胞,诱导转基因神经干细胞贴壁分化,观察其体外增殖、分化特点。采用RT-PCR技术检测干细胞和分化细胞表面标志或相关基因。【结果】成功分离培养出神经干细胞,获得转EGFP基因神经干细胞,神经干细胞在表达EGFP的同时仍具有多向分化潜能。神经干细胞中Nestin表达强阳性,NogoA、DCX、CyclinD2、CD133、Hes1、Oct4、CD-90、Nanog和Sox2表达阳性;体外诱导的神经干细胞可以分化为星形胶质细胞(表达GFAP)、少突胶质细胞(表达GalC)和神经元细胞(表达NF、NSE和MAP2);能分化为脂肪细胞(表达LPL和PPARγ-D)、成骨细胞(表达Osteonectin和Osteocalcin)、肌细胞(表达myf-5、myf-6和myoD)、内皮细胞(表达CD31、CD34、CD144和eNOS)和软骨细胞(表达COL2A1)。【结论】从猪胎儿大脑组织分离神经干细胞具有可行性和有效性,转EGFP基因神经干细胞具有自我更新、增殖和多向分化潜能,可以用EGFP对神经干细胞进行标记、追踪,作为示踪标记进行神经干细胞体内移植研究。     

云南地方香稻与非香稻遗传多样性比较

[目的]比较云南地方香稻品种与非香稻地方栽培品种的遗传多样性。[方法]利用分布于水稻12条染色体上的64对SSR引物检测云南的10个香稻品种、45个非香稻地方栽培品种。[结果]在云南香稻资源中,每对引物检测到的等位基因数为2-12个,平均每个位点的等位基因数为5.44个,平均基因多样性（Hs）和平均多态信息含量（PIC）分别为0.46和0.43;而在云南非香稻栽培品种中,每对引物检测到等位基因数2-17个,平均每个位点的等位基因数为7.98个,平均基因多样性（Hs）和平均多态信息含量（PIC）为0.67和0.58。[结论]云南非香地方栽培稻资源比香稻资源具有更为丰富的遗传多样性。     

Identification of molecular markers associated with sugar-related traits in a <Emphasis Type="Italic">Saccharum</Emphasis> interspecific cross

The cultivated sugarcane (Saccharum spp. hybrids, 2n = 100–130) is one crop for which interspecific hybridization involving wild germplasm has provided a major breakthrough in its improvement. Few clones were used in the initial hybridization event leading to a narrow genetic base for continued cultivar development. Molecular breeding would facilitate the identification and introgression of novel alleles/genes from the wild germplasm into cultivated sugarcane. We report the identification of molecular markers associated with sugar-related traits using an F₁ population derived from a cross between S. officinarum ‘Louisiana Striped’ × S. spontaneum ‘SES 147B’, the two major progenitor species of cultivated sugarcane. Genetic linkage maps of the S. officinarum and S. spontaneum parents were produced using the AFLP, SRAP and TRAP molecular marker techniques. The mapping population was evaluated for sugar-related traits namely, Brix (B) and pol (P) at the early (E) and late (L) plant growing season in the plant cane (04) and first ratoon (05) crops (04EB, 04LB, 04LP, 05EB and 05EP). For S. officinarum, combined across all the traits, a total of 30 putative QTLs was observed with LOD scores ranging from 2.51 to 7.48. The phenotypic variation (adj. R²) explained by all QTLs per trait ranged from 22.1% (04LP) to 48.4% (04EB). For S. spontaneum, a total of 11 putative QTLs was observed with LOD scores ranging from 2.62 to 4.70 and adj. R² ranging from 9.3% (04LP) to 43.0% (04LB). Nine digenic interactions (iQTL) were observed in S. officinarum whereas only three were observed in S. spontaneum. About half of the QTLs contributed by both progenitor species were associated with effects on the trait that was contrary to expectations based on the phenotype of the parent contributing the allele. Quantitative trait loci and their associated effects were consistent across crop-years and growing seasons with very few QTLs being unique to the early season. When the data were reanalyzed using the non-parametric discriminant analysis (DA) approach, significant marker-trait associations were detected for markers that were either identical to or in the vicinity of markers previously identified using the traditional QTL approach. Discriminant analysis also pointed to previously unidentified markers some of which remained unlinked on the map. These preliminary results suggest that DA could be used as a complementary approach to traditional QTL analysis in a crop like sugarcane for which saturated linkage maps are unavailable or difficult to obtain.     

Genetic stability at nuclear and plastid DNA level in regenerated plants of <Emphasis Type="Italic">Solanum</Emphasis> species and hybrids

In this work we detected the extent of variability at nuclear and cytoplasmic DNA level of regenerated plants belonging to Solanum genotypes with a different genetic background and somatic chromosome number. As for the nuclear characterization, a total of 66 (18.5%) polymorphic bands were scored using 13 ISSR primers on 45 randomly selected regenerants. Our results show that the regenerants obtained from clone cmm 1T and, at lower level, those from cph 1C are unstable under in vitro conditions or rather more prone to in vitro-induced stress leading to somaclonal variation than the other genotypes used. Two types of changes were observed: disappearance of parental ISSR fragments, termed “loss”; appearance of novel ISSR fragments, termed “gain”. The most frequent event occurring in the regenerants was the loss of fragments (41 bands). Regenerated plants were analyzed with seven plastid universal primers to determine the cytoplasmic composition at chloroplast level. All cpDNA primer pairs tested produced amplicons of the same size in all genotypes analyzed and no polymorphic fragments were observed with any universal primers used. Our results show that under in vitro culture conditions genotype affects the integrity of the genome. In addition, the absence of polymorphism at plastid level confirms the greater genetic stability of cytoplasmic DNA.