从干种子中快速获取高质量DNA的高盐提取方法

为寻求一种从植物干种子中简单快速提取高质量基因组DNA的有效方法进行本研究。将种子打磨成粉,取100 mg粉末加入高盐提取液抽提基因组DNA。采用超微量分光光度计检测、PCR及酶切的方法检验DNA的得率和质量。从100 mg 7种常见作物种子粉末中可以得到619.67~1811.21 ng基因组DNA,A260/A280比值在1.87~2.07之间,其纯度和质量适合进行PCR及酶切分析。通过普通PCR方法分别对7种作物的特异性内源基因片段进行扩增,结果均能扩增到明显的目的条带。用EcoRV和HindIII两种核酸内切酶能对所得的DNA充分酶切。结果表明本方法能快速地从干种子中提取到高质量的DNA。     

利用RAMP-PCR技术对枸杞10个品种资源的分析

以枸杞为材料,建立基因组DNA的有效提取方法,并对10个枸杞品种进行了RAMP技术分析研究。结果表明,利用10对引物共扩增出87个条带,其中多态性条带53个;多态性条带比率33.3%~81.9%,10个枸杞品种间的遗传距离为0.1111~0.8333;研究表明,利用RAMP进行枸杞的品种DNA指纹分析是可行的。     

不同作物品种对60Co γ射线的辐照敏感性

为了了解不同作物品种间对⁶⁰Co γ射线的辐照敏感性,作者用0~1500Gy ⁶⁰Co γ 辐射剂量对棉花、水稻、大麦、油菜、大豆和玉米等作物7个品种的种子进行了辐照。结果显示发芽率呈现：（1）不敏感的油菜;（2）随辐照剂量的增加、发芽率逐渐下降的水稻和大豆;（3）发芽率呈“下降-上升-再下降”的棉花和大麦,但最高发芽率未超过对照;和（4）发芽率出现先上升后下降,辐照剂量为300Gy 时发芽率达到最大值的糯玉米和甜玉米等4种类型的变化。辐照敏感性依次为水稻>大豆>棉花>糯玉米、甜玉米>大麦>油菜。因此,作者认为不同作物辐照敏感性的差异除了油菜种子因内含对辐射有屏障作用的丙烯芥子油外,主要受作物基因组大小和种子饱满度的影响。     

侵染白菜的黄瓜花叶病毒分离物基因组的全序列分析

对浙江省杭州地区白菜（Brassica campestris L. ssp. chinensis var. commuis Tsen et Lee.）上获得的CMV分离物（CMV-CTL）进行了全长克隆和基因组序列分析。结果显示：CMV-CTL的RNA1（GenBank序列号：EF213023）全长为3357个核苷酸（nt）,编码993个氨基酸（aa）的1a蛋白;RNA2 （GenBank序列号：EF213024）全长为3047 nt ,编码858 aa的2a蛋白和111 aa的2b蛋白;RNA3 （GenBank序列号：EF213025）全长为2217 nt,编码278 aa的MP蛋白和218 aa的CP蛋白。序列相似性分析表明,CMV-CTL与CMV亚组IB中株系IA相似性最高,RNA 1、RNA 2和RNA3与该株系的相似性分别为91.3%、91.3%和93.6%。CP基因和RNA 3 的5' NTR核酸序列系统发生树分析表明,CMV-CTL与中国大多数CMV分离物一样,属于CMV亚组IB。     

Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

Haynaldia villosa (2n =2X = 14, VV), a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetie Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge (a remarkable morphology) mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V(2D) and the hybrid F1 was then self-crossed. Chromosome C-banding, genomie in situ hybridization (GISH), and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids F2 and F3. To date, four translocations including one small segmental translocation T6BS.6BL-2VS, two whole arm translocations (preliminarily designed as T3DS·2VL and T2VS·7DL) and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one iso- chromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25-120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 (2BL) tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS·7DL, Mt2VS, 2VS·2VS as well as the parent DS2V(2D), whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C of Aegilops triuncialis could successfully induce chromosome 2V structural changes.     

转小粒野生稻基因组DNA水稻的产量与品质性状研究

野威B是将小粒野生稻(Oryza minuta)的基因组DNA通过穗茎注射法导入杂交水稻亲本V20B中培育出的转基因水稻新种质.与亲本V20B相比,野威B的有效穗数和每穗总粒数小于V20B,但每穗实粒数却高于V20B,平均结实率比V20B增加11.8%,千粒重比V20B的少6 g.野威B倒1叶和倒2叶叶鞘中可溶性糖含量...     

Conventional and molecular diagnostic testing for the acute neurologic patient

Objective – The aim of this review is to describe and evaluate both conventional and molecular diagnostic testing utilized in dogs and cats with acute neurologic diseases. Various types of polymerase chain reaction (PCR) are explored along with novel molecular diagnostic testing that ultimately may prove useful in the critical care setting. Data Sources – PUBMED was searched to obtain relevant references material using keywords: ‘canine OR feline meningitis AND meningoencephalitis,’‘feline infectious peritonitis,’‘canine distemper,’‘canine OR feline AND toxoplasma,’‘canine neospora,’‘canine OR feline AND rickettsia,’‘granulomatous meningoencephalitis,’‘steroid responsive meningitis arteritis,’‘necrotizing encephalitis,’‘novel neurodiagnostics,’‘canine OR feline AND CNS borrelia,’‘canine OR feline AND CNS bartonella,’‘canine OR feline AND CNS fungal,’‘nested OR multiplex OR degenerate OR consensus OR CODEHOP AND PCR.’ Research findings from the authors' laboratory and current veterinary textbooks also were utilized. Human Data Synthesis – Molecular diagnostic testing including conventional, real‐time, and consensus and degenerate PCR and microarray analysis are utilized routinely for the antemortem diagnosis of infectious meningoencephalitis (ME) in humans. Recently, PCR using consensus degenerate hybrid primers (CODEHOP) has been used to identify and characterize a number of novel human viruses. Veterinary Data Synthesis – Molecular diagnostic testing such as conventional and real‐time PCR aid in the diagnosis of several important central nervous system infectious agents including canine distemper virus, Toxoplasma gondii, Neospora caninum, rickettsial species, and others. Recently, broadly reactive consensus and degenerate PCR reactions have been applied to canine ME including assays for rickettsial organisms, Borrelia spp. and Bartonella spp., and various viral families. Conclusions – In the acute neurologic patient, there are several key infectious diseases that can be pursued by a combination of conventional and molecular diagnostic testing. It is important that the clinician understands the utility, as well as the limitations, of the various neurodiagnostic tests that are available.     

利用栽培稻基因组DNA对非洲野生稻进行染色体荧光原位杂交分析

以地高辛标记的栽培稻基因组(基因组为AA)DNA为探针,对非洲野生稻(基因组为BBCC)的体细胞染色体进行荧光原位杂交分析,研究AA染色体组和BBCC染色体组之间的关系,同时对杂交后的染色体进行同源染色体配对。结果表明:栽培稻A基因组和非洲野生稻基因组有较高的同源性,其中高度重复DNA序列在栽培稻和非洲野生稻间具有保守性。     

小麦成熟胚脱分化过程中信号转导相关基因的初步分析

 【目的】研究信号转导相关基因在小麦成熟胚脱分化过程中的表达模式以揭示其脱分化的分子机理。【方法】利用Affymetrix小麦基因芯片研究了小麦成熟胚在MS+2,4-D（2 mg•L1）培养基上脱分化过程不同时间点的基因表达变化,通过NCBI、DATF和DRTF等生物信息学相关网站对基因表达信息进行处理,对信号转导相关基因的变化情况进行了分析,用半定量RT-PCR方法对AF161719.1、BE492852、BJ218430、BJ252470、BJ272518、BJ274015、CA616149、CD896892、CK153462、CK207725、CK171662、CD901339和U48692.1基因进行验证。【结果】有46个信号转导相关基因在小麦成熟胚脱分化过程中的2、6、12、24和72 h等不同时间点至少发生了一次有意义的表达变化,其中在整个过程中有32个基因上调,14个基因下调。这些基因包括酪蛋白基因、苏氨酸丝氨酸蛋白激酶基因、14-3-3蛋白基因、钙调素基因和裂原激活蛋白激酶基因,其表达变化与基因芯片的结果基本一致。【结论】CA613597、CA674315、BJ274015、CA654806、CD901339、CK198900、CA697195、CA642143、CA744550和AB011670.1对脱分化的启动和促进可能起重要作用。