芸薹属A基因组DNA封阻下的甘蓝C染色体组核型分析

为探索一种高效可靠的芸薹属植物核型分析方法,以甘蓝(Brassica oleracea)和白菜(B.pekinensis)为材料,提取甘蓝基因组DNA用地高辛标记为探针,提取白菜基因组DNA作为封阻剂,对甘蓝有丝分裂中期染色体进行原位杂交.结果表明,甘蓝每对同源染色体上均显示出了特定的原位杂交信号,依据信号特征成功地将甘蓝9对染色体进行了精细的分型.为甘蓝等小型染色体的核型分析提供了一条可行且精确的方法.     

静宁葫芦巴染色体制片及核型分析

以采自静宁的葫芦巴为材料,研究了1 mol/L的盐酸在60 ℃下的不同解离时间对根尖染色体制片的影响,并确定染色体数目和进行核型分析。结果表明,静宁葫芦巴根尖最佳解离时间为2.0～2.5 min。染色体数为16,核型公式为2n＝2x=16=8m+8sm(2SAT),臂比值变化在1.07～2.88,平均臂比为1.926,随体在第6对染色体短臂上,核型类别为2A。     

3种不同皮色萝卜种质染色体核型分析

本研究采用根尖压片法,对3种不同肉质根皮色的萝卜种质进行染色体数目和核型分析,以获得准确的细胞遗传学信息。研究结果表明,Nau-dqp08,Nau-txbyw07和Nau-chhong108这3种不同皮色萝卜的染色体数目均为18,核型公式分别为2n=2×=18=14m+4sm,2n=2×=18=18m和2n=2×=18=16m+2sm,且核型都属于1A型;核不对称系数分别为59.48%、55.39%与58.08%;核型不对称性大小为Nau-dqp08>Nau-chhong108>Nau-txbyw07,其中Nau-txbyw07核型较为原始。本研究结果可为萝卜种质鉴定、遗传变异和亲缘关系分析提供细胞学依据。     

入侵杂草假高粱的染色体变异及核型分析

对入侵杂草假高粱(Sorghum halepenseL.)的根尖细胞进行染色体计数及核型分析,同时与二倍体高粱(SorghumL.)作比较。结果表明:供试材料假高粱的染色体数目为2n=34,染色体基数x=17,与原产地同物种(x=10)相比,发生了染色体数目变异,其核型公式为2n=2x=34=24m(2SAT)+10sm,为异源四倍体植物,核型类型为2B型,第15号染色体上有一对随体,最长与最短染色体长度比为2.30,臂比>2的染色体比例为18%,核型不对称系数为60.02%。与核型为2A型的高粱相比,二者的染色体数不成倍性关系,表明该假高粱种可能是由入侵种假高粱和禾本科其他植物通过远缘杂交形成的新变种。     

滁菊染色体核型的微变异

以安徽滁州地区特有的药食两用植物资源——滁菊(Dendranthema morifolium Ramat.‘Chuju’)为试材,采用根尖细胞压片技术进行染色体计数及核型分析,并与引种到外地的滁菊种进行比较。结果表明:滁菊种的染色体数目为2n=54,染色体基数x=9,为异源六倍体植物,其核型公式为2n=6x=54=42m+12sm(4SAT),核型类型为2A型,第10和21号染色体上均有1对随体,最长与最短染色体长度比为1.99,臂比>2的染色体比例为18.51%。与已公布的引种到外地的滁菊核型进行比较分析发现,引种后的滁菊较原生境滁菊染色体核型发生了微变异,表现在具随体染色体数量由4条减为2条,染色体多态性更为丰富,提示滁菊染色体核型与其生态环境有着密切的关系。     

小叶丹参的核型分析

以小叶丹参(Salvia miltiorrhiza Bge)试管苗茎尖为材料,采用压片法获得2倍体染色体2n=16.其核型公式为2n=2x=14m 2sm=16.     

适用于RAPD分析的蚕卵基因组DNA快速提取方法的探讨

目前所报道的家蚕基因组DNA的提取方法(Sharp et a1,1988;Yang et al,1993;夏庆友等,1996;王韵等,1997)基本上都是先应用离子型表面活性剂(SDS)及蛋白酶K去除结合在核酸上的蛋白质,然后用酚、氯仿变性除去蛋白,RNase去除RNA,最后乙醇沉淀DNA获得.这些方法操作过程繁复、耗时多,需用酚、氯仿等有毒试剂.本实验尝试进行改良,寻找适用于大量样品作RAPD分析并简单、快捷、低成本的蚕卵基因组DNA的提取方法.     

薄片牡蛎的核型及18S-28S核糖体rRNA基因的染色体定位

以薄片牡蛎（Dendostrea folium）成体鳃组织为材料制备有丝分裂中期染色体标本,对其染色体核型进行了分析,并运用荧光原位杂交技术（FISH）将18S-28S核糖体RNA基因定位于中期染色体上。FISH探针是通过PCR扩增介于18S-28S rRNA基因之间的ITS和5.8S rRNA基因序列,并在PCR扩增过程中掺入了Biotin-11-dUTP进行生物素标记。结果显示,薄片牡蛎的单倍染色体数目为n=10,全部为中部着丝粒染色体。与大多数已知巨蛎属牡蛎的染色体核型相似。ITS探针在薄片牡蛎中期分裂体相上产生两簇FISH信号,分别杂交于2号染色体短臂的近端粒区域。本研究首次报道了薄片牡蛎的中期染色体核型以及18S-28S核糖体RNA基因在染色体上的定位。     

鸡脚重性状的全基因组关联分析

鸡(Gallus gallus)脚重性状为鸡产业中的重要经济性状.为了揭示鸡脚重性状的遗传基础,寻找可用于鸡脚改良的分子标记,本研究以核心群京海黄鸡为实验动物,测定其脚重,利用简化基因组测序技术在整个基因组范围内检测与脚重相关的SNPs.共检测到16个与脚重相关的SNPs位点,其中13个集中分布在4号染色体上72.8～77.9 Mb区域;2个SNP位点rs475641139和rs475548810达到5％ Bonferroni全基因组显著水平(P＜5.5E-07),其余位点达到全基因组潜在显著水平(P＜1.1E-05).筛选每个SNP周围1 Mb区域内的基因,共找到9个可能的候选基因,并使用基因本体论(Gene Ontology,GO)数据库对9个候选基因的细胞组分、分子功能和参与的生物学进程进行了分析.结果表明二氢蝶啶还原酶(dihydropteridine deductase,QDPR)基因、LIM域结合蛋白2(LIM domain-binding protein 2,LDB2)基因、类184B家族(family with sequence similarity 184 memberB,FAM184B)基因、复合信号免疫球蛋白J结合蛋白(recombination signal binding protein for immunoglobulin kappa J region,RBPJ)基因、纤维母细胞生长因子结合蛋白2(fibroblast growth factor-binding protein 2,FGFBP2)基因、富亮氨酸重复蛋白5(F-box and leucine-rich repeat protein 5,FBXL5)基因、胆囊收缩素受体A(cholecystokinin receptor type A,CCK1R)基因、肌动蛋白互作蛋白1 (WD repeat containing protein 1,WDR1)基因和类桶状蛋白4(tubby like protein 4,TULP4)9个基因和4号染色体72.8～77.9 Mb区域可能为影响鸡脚重性状的重要候选基因和潜在功能区域.本研究为鸡脚重性状的标记辅助选择提供了理论依据.     

斜纹夜蛾核多角体病毒基因组DNA XbaI 2.0 kb片段全序列分析

报道了斜纹夜蛾核多角体病毒（Ｓｐｏｄｏｐｔｅｒａ ｌｉｔｕｒａ ｎｕｃｌｅｏｐｏｌｙｈｅｄｒｏｖｉｒｕｓ,ＳｐｌｔＮＰＶ）基因组ＤＮＡ ＸｂａＩ２．０ｋｂ片段的核苷酸全序列,该片段包括三个完整的读码框（ｏｐｅｎ ｒｅａｄｉｎｇ ｆｒａｍｅ,ＯＲＦ）,即ＯＲＦ１、ＯＲＦ２、ＯＲＦ３及一个不完整的读码框内氨基酸有１８％相同;ＯＲＦ２长５７０个核苷酸,可编码１８９个氨基酸的蛋白质,分子量为２１．６４ｋＤ     

Phylogenetic relationships among Triticum L. and Aegilops L. species as genome progenitors of bread wheat based on sequence diversity in trnT-F region of chloroplast DNA

Cultivated wheat, (Triticum aestivum L.), is one of the most important food crops in the world. The Aegilops L. genus is frequently utilized by plant breeders for improving the current wheat cultivars due to their close relationships. Therefore, understanding the phylogenetic relationships among the species of these genera is not only valuable for plant taxonomy, but also for plant breeding efforts. The presented phylogenetic analysis was based on the sequences of trnT-F chloroplast DNA containing three non-coding sub-regions. Twelve genotypes belonging to four species of Triticum L. genus and twenty-four genotypes belonging to eight species of Aegilops genus were used in the current study. The results postulated a close genetic relationship between diploid Aegilops species containing the BB genome and polyploid Triticum species. With the exception of Aegilops cylindrica Host (CCDD), all other Aegilops species having the CC genome were alienated from Aegilops speltoides Tausch (BB) and clustered together. These two clusters joined by a third cluster including the AA genome containing diploid Triticum species.     

Trait inheritance,fertility, and genomic relationships of some n = 9 Brassica species

Summary Karyotypic differences among n = 9 Brassica species, as well as the inheritance of some morphological characteristics, are reported. Annual habit, anthocyanin pigmentation in leaf and stem, enlarged stem, glossy foliage, dark green foliage, white flower and leaf pubescence were all expressed as dominant characteristics and in most cases were determined by one or two genes. 172 intra- and interspecific hybrids of the species belonging to the Brassica oleracea cytodeme were analyzed for fertility and meiotic chromosome behavior. B. alboglabra, B. bourgeaui, B. cretica spp. cretica, B. montana and B. oleracea produced fertile interspecific hybrids, indicating that geographical isolation is the only barrier to gene exchange. Interspecific hybrids generated from crosses of B. incana, B. insularis and B. rupestris to other n = 9 species were semi-sterile. This was shown to be associated with abnormal meiotic behavior. The former two species, when crossed, produced fully fertile hybrids. Therefore, three different karyotypes are identified among the n = 9 Brassica species. Frequent changes in the genome of the species belonging to the Brassica oleracea cytodeme could have led to the formation of accessory chromosomes present in certain accessions.     

Cytoplasmic diversity of Brassica napus L., Brassica oleracea L. and Brassica rapa L. as determined by chloroplast microsatellite markers

Cytoplasmic genomes in most angiosperms are known to be maternally inherited. Oilseed rape (Brassica napus L.) as a natural amphidiploid species hence may carry the B. oleracea L. or the B. rapa L. cytoplasm, depending on the cross direction. The presence of either the B. oleracea or the B. rapa cytoplasm in oilseed rape has been reported to affect agronomically important traits. However, to date little is known about the cytoplasmic composition and genetic diversity of current winter oilseed rape cultivars and breeding material. The aim of this study was to assess the usefulness of 40 previously published chloroplast cpSSR markers from Brassica species and Arabidopsis thaliana (L.) Heynh. for distinguishing the cytoplasms of 49 different genotypes of B. napus and its diploid ancestor species. Results showed that only 14 out of the 40 tested primer combinations were suitable to distinguish the cytoplasms of a test set of 8 Brassica genotypes. With the 14 primer pairs 64 different cpSSR alleles were identified in the set of 49 genotypes. Cluster analysis indicated distinct groups for the cytoplasms of B. napus, B. rapa, and B. oleracea. However, an unambiguous identification and classification of the cytoplasm types was not possible in all cases with the available polymorphic set of cpSSR primer pairs.     

基于块标记的田间叶片损伤区域分割方法

为了在田间开放环境中有效分割叶片损伤区域,该文结合Canny算子良好的边缘提取能力和叶片局部颜色变化相对较小的特征,提出基于块标记的叶片损伤区域分割方法,用于评价叶片损伤程度。使用Android系统手机在晴天大田开放环境中采集木耳菜、西红柿、黄瓜、茄子、桃、彩椒和蛾眉豆7种常见农作物叶片图像,在阴天采集丝瓜、葫芦、甜瓜、茄子和黄瓜5种叶片图像,然后进行分割。该分割算法在晴天和阴天总体的平均正确分类率为97.5%,平均错误分类率为0.3%,并且有较好的目标一致性和边缘清晰度。应用系统对叶片损伤程度的评价结果与手工分割比较,在晴天和阴天采集图像上的平均误差分别为2.340%和1.475%,可较好地应用于晴天和阴天环境。该方法可探索应用于田间植物叶片损伤程度评价。     

碱性土壤基因组DNA的分离纯化和基因文库的构建

直接从广西南宁市凤凰纸业排污沟碱性土壤样品中抽提和分离纯化混合基因组DNA，所获得DNA的产量为每克土壤样品10～30pg。采用限制性内切酶EcoR1酶处理后，构建了以pGEM-3Zf( )为载体的DNA部分文库。文库的容量为23650个转化子。外源片段DNA平均大小为3．2kb。建库效率为每克环境样品获得6000个左右的含1～15kb外源随机插入片段的克隆。通过DNA序列测定和同源性比较，对从文库随机词取的16个转化子序列进行分析，发现13个外源插入片段包含序列尚未确定的DNA片段。     

The origin of the A genome donor of wheats (Triticum: Poaceae) – a perspective based on the sequence variation of the 5S DNA gene units

In a prior study on the haplomes of wheat using the 5S rRNA gene we assigned the long A1 and short A1 unit classes to the A haplome in the diploid T. monococcum. The short A1 unit class is absent in the tetraploids T. turgidum and T. timopheevii and in the hexaploid T. aestivum, although present in the hexaploid T. zhukovskyi. Both T. turgidum and T. aestivum contained a different 5S DNA unit class labeled the short A2.The purpose of this paper was to study the short A2 units in the two diploid species to shed light on the theory that the A haplome donor of T. turgidum and T. aestivum was T. urartu. Fifty eight clones were obtained from 12 accessions, sequenced and analyzed. As expected T. baeoticum, which is often classified as a subspecies of T. monococcum, contained the long A1 and the short A1 5S DNA units. Unexpectedly, T. urartu had the long A1 and the short G1 unit classes instead and other units not found so far in Triticum. These findings support the hypothesis that the donor of the A genome in T. zhukovskyi was T. monococcum, as identified by the short A1 units. However, the short A1 units are absent in T. timopheevii, also a carrier of the A genome. The short G1 units found in T. urartu also identify it as a possible donor of the G genome to T. timopheevii. The short G1 units were also found in T. aestivum in our prior study. The long G1 unit class was not found in T. urartu but reported from T. timopheevii and T. zhukovskyi. The implications of these and related findings on the evolution of wheats are discussed.     

The role of EDTA in malonaldehyde formation from DNA oxidized by Fenton reagent systems

Calf thymus DNA was oxidized by various Fenton reagent systems [Fe(II)/H(2)O(2)] with or without ethylenediamine tetraacetic acid (EDTA) under different reaction conditions. Calf DNA was also oxidized by a modified Fenton reagent (Fe(III)/H(2)O(2)/ascorbic acid) with EDTA. Malonaldehyde (MA) formed from DNA was derivatized into 1-methyl hydrazine, which was subsequently analyzed by gas chromatography with a nitrogen-phosphorus detector. MA formation increased linearly with an increase of Fe(II) concentration. MA formation reached a plateau at nearly 2 mmol/L of Fe(II) with 0.5 mmol/L of H(2)O(2). Addition of EDTA increased MA formation from DNA nearly 5 times. When DNA was oxidized with various amount of ethanol, MA formation decreased with an increase of ethanol concentration, either with or without EDTA. The rate of inhibition was greater without EDTA than with EDTA. When DNA was oxidized by a modified Fenton reagent, MA formation linearly increased with the increase of DNA. Ascorbic acid alone produced some MA upon oxidation.     

Evaluation of Mehlich 3 reagent as a multielement extractant in mine soils

The usefulness of Mehlich 3 (M3) reagent was evaluated as a method to extract numerous elements from coalmine soils in As Pontes (Spain) showing a wide range of physicochemical properties. Critical levels (deficiency and/or toxicity) were established for plant available elements extracted by this reagent. The M3 method was compared to 1M NH₄Cl, Olsen, acid oxalate, and DTPA methods as extractants for exchangeable Ca, Mg and K, available P, non-crystalline aluminium, and available heavy metals, respectively. The M3 method correlated significantly to NH₄Cl for Ca, Mg and K (r=0·76, 0·84 and 0·87, respectively), to Olsen P (r=0·77) and to oxalate Al (r=0·77). Significant correlations were found between Fe, Cu, Zn and Cd extracted by M3 and DTPA; for Mn, Ni, Co and Pb different relationship between methods were obtained for acid and alkaline samples, so that critical levels were established for M3 metals as a function of soil pH. Copyright © 1999 John Wiley & Sons, Ltd.