敲减MC4R表达对牛胎儿成纤维细胞CMS系统关键因子的影响

为获得敲减黑素皮质素4受体（melanocortin 4 receptor,MC4R）基因的牛胎儿成纤维细胞,并探讨其在能量平衡神经调节系统中的作用,将构建成功并已鉴定为有效序列的短发夹状RNA (short hairpin RNA, shRNA)真核表达载体pGSH1 GFP MC4R,利用阳离子脂质体转染牛胎儿成纤维细胞并使用G418筛选稳定转染细胞株.利用实时荧光定量和Western印迹检测MC4R及中枢黑素皮质素系统(central melanocortin system, CMS)关键因子的表达水平变化.结果表明,在稳定转染的牛胎儿成纤维细胞系中, MC4R表达显著抑制,瘦蛋白(leptin)和阿黑色素原(POMC)表达下调,黑素皮质素拮抗物agouti相关蛋白(AGRP)和MC3R表达上调,而神经肽Y (NPY)表达无明显改变.综上所述,本研究成功获得了敲减MC4R基因表达的牛胎儿成纤维细胞.相关基因表达水平检测结果提示, MC4R的表达水平对CMS系统中的各关键基因的表达有不同的抑制或促进影响.     

基于BSA-seq法的油菜野芥胞质雄性不育恢复基因的分析

细胞质雄性不育是利用杂种优势创制油菜杂交种的重要途径之一。而恢复系的选育及恢复基因的研究有利于提高细胞质雄性不育系的应用价值。该研究以甘蓝型油菜细胞质雄性不育系1193A和恢复系15-R1为亲本,以F_2群体构建了2个极端性状混池,采用BSA-seq技术方法,对恢复基因进行定位分析。结果表明:(1)不育系1193A与恢复系15-R1之间共获得非同义突变的SNPs共33 883个,混池间共有7 996个非同义编码突变。(2)InDel检测与注释结果表明,亲本间引起移码突变的有2 918个,混池间引起移码突变的有840个。(3)综合SNP和InDel的关联分析结果,将恢复基因定位于C09染色体上的0～880000区域,总长度为880 kb。(4)对候选区域的SNP和InDel注释发现,亲本间存在非同义突变的SNP共40个,混池间存在非同义突变的SNP共65个,亲本间存在移码突变的InDel共7个,混池间存在移码突变的InDel共11个;对候选区间内的编码基因进行注释,结果共注释到162个基因,其中存在非同义突变基因11个,移码突变基因5个,这些基因可能与育性恢复有关。     

Aspects of interspecific hybridization within edible <Emphasis Type="Italic">Alliaceae</Emphasis>

Crossing experiments within the genus Allium have been carried out for many years. Usually, with the aim of widening genetic variation, edible Alliaceae such as bulb onion, Japanese bunching onion, leek, garlic and shallot have been crossed with each other or with distant vegetable Allium crops. Interspecific hybridization, especially with wild relatives is considered as the best way of introgression desirable traits to the crops. By using sexual or somatic hybridization, many important characters, such as: disease and pest resistance, important metabolites and cytoplasmic male sterility have been transferred to other Allium crops. This review summarises some aspects of the directions of interspecific hybridization in edible Alliaceae and the significance and perspectives of using these hybrids in breeding programmes.     

特种稻不育系巨胚813A的选育与研究

用350Gy剂量的Co^60-γ射线照射水稻三系保持系813B种子,获得巨胚突变体,命名为巨胚813B（简称813geB）。用巨胚813B回交813A育成特种稻不育系——巨胚813A（简称813geA）。对巨胚813B及巨胚813A的生物学特性进行研究,结果表明：巨胚813A与813A的主要农艺性状、雄性不育性及产量构成因素均无明显差异。巨胚813B与813B具有相似的农艺性状,但绝对胚重由0．61mg提高到1．36mg,相对胚重由2．70％提高到6．96％。巨胚813B糙米的蛋白质、粗脂肪、粗纤维及必需氨基酸含量均比813B高。其中蛋白质含量9．77％,比813B提高了8．80％;粗脂肪含量5．28％,比813B提高了87．23％;8种必需氨基酸含量均有提高。利用巨胚恢复系与巨胚813A可纽配成巨胚杂交稻。     

利用野生青狗尾草的细胞质培育谷子质核互作雄性不育材料

利用野生青狗尾草N10为母本,谷子农家品种大青秸为父本,进行种间杂交,获得了3株杂种。杂种在农艺性状上表现为谷子和青狗尾草的种间类型,花器表现了雄性败育,但雌蕊发育正常,能接受外来花粉结实。细胞形态学观察表明,其雄性败育表现为单核小孢子典败。该谷子雄性不育材料的获得,为利用野生青狗尾草的细胞质培育谷子质核互作雄性不育系,进而为实现谷子三系配套杂种优势利用奠定了基础。     

六倍体小黑麦T型细胞质雄性不育体系杂种优势与配合力的研究

用具提莫菲维小麦细胞质的六倍体小黑麦的3个不育系和3个恢复系作为亲本,进行3×3不完全双列杂交,对所组配的F1代8个农艺性状的杂种优势分析结果表明,除千粒重外,其余性状出现正向超亲优势的组合较少,多数呈低亲或中亲遗传,且各组合间的差异比较显著。配合力分析表明,一般配合力与特殊配合力的方差均达到了显著水平,F1各性状均受基因加性效应和非加性效应共同作用;从总体上看,不育系A1、A2及恢复系R1、R2的一般配合力良好,其配制的组合优势较强,具一定的利用价值。对一般配合力与亲本表型值进行了相关分析,二者无显著的相关关系。     

DNA methylation changes in photoperiod-thermo-sensitive male sterile rice PA64S under two different conditions

Epigenetic modification can occur at a high frequency in crop plants and might generate phenotypic variation without changes in DNA sequences. DNA methylation is an important epigenetic modification that may contribute to environmentally-induced phenotypic variations by regulating gene expression. Rice Photoperiod-Thermo-Sensitive Genic Male Sterile (PTGMS) lines can transform from sterility to fertility under lower temperatures and short-day (SD) conditions during anther development. So far, little is known about the DNA methylation variation of PTGMS throughout the genome in rice. In this study, we investigated DNA cytosine methylation alterations in the young panicles of PTGMS line PA64S under two different conditions using methylation sensitive amplified polymorphism (MSAP) method. Compared with the DNA methylation level of PA64S under lower temperatures and SD conditions (fertility), higher methylation was observed in PA64S (sterility). The sequences of 25 differentially amplified fragments were successfully obtained and annotated. Three methylated fragments, which are homologous to D2, NAD7 and psaA, were confirmed by bisulfite sequencing and their expression levels were also evaluated by qPCR. Real time quantitative PCR analysis revealed that five of the six selected methylated genes were downregulated in PA64S (sterility). These results suggested that DNA methylation may be involved in the sterility–fertility transition of PA64S under two different environmental conditions.     

结球白菜温度敏感细胞质雄性不育系CMS7311育性转换的研究

在人工控温和田间自然变温条件下,研究了结球白菜9Brassica campestris L.ssp.penkinsis)胞质雄性不育系CMS7311育性转换的温度特征。结果表明：随温度变化CMS7311有明显的育性转换,人工恒温和人工变温条件下诱导CMS7311发生育性转换（不育→可育）的温度范围为日均温6－12℃,有效诱导的最短时间为3d,显诱导的温度范围为6－9℃。自然变温条件下,有效诱导的温度低限为日均温3.5-6.8℃,但诱导育性转换的最短时间要适当加长。无论恒温还是变温,在日均温9℃下处理9d育性转换达到最大,接近正常可育（0级或1级）。正常情况下,温度处理后到发生育性转换大约需要10－16d,育性转换发生在孢原细胞形成至花粉分化阶段。由于白菜生长的顶端优势作用,在相同诱导条件下,同一植株侧枝较主枝的育性转换时间晚8－10d,且转换程度较弱。     

Some physicochemical properties of rice mitochondrial DNA

Summary Certain physicochemical properties of rice mitochondrial DNA (mtDNA) were determined. Certain low-molecular-weight mtDNA bands were found in addition to the major mtDNA band. Rice mtDNA appeared in the electron microscope as a collection of linear molecules with heterogeneous length in the range of 1–156 kb. The major distribution area was 60–105 kb. A small fraction (less than 5%) of rice mtDNA was found in the form of a circular molecule. Some molecules had the appearance of being supercoiled. Replication fork structures were found in both circular and linear mtDNA molecules. In one rice species, Jin Nante, 15 different circular molecules were found. Rice mtDNA was digested with different restriction enzymes. The total molecular weight of rice mtDNA was calculated to be about 300 kb according to the data of restriction enzyme digestion and electron microscopy.