应用柑橘异源四倍体杂种花粉生产瘪籽沙田柚果实

以3个柑橘异源四倍体体细胞杂种(即四倍体粗柠檬与哈姆林甜橙体细胞杂种,简称"HR";酸柚与粗柠檬体细胞杂种,简称"SR";墨西哥来檬与伏令夏甜橙体细胞杂种,简称"KV")为父本,分别与二倍体单胚性沙田柚进行有性杂交,在生产上获得瘪籽沙田柚果实,并对果实品质进行分析(以四季柚花粉亲本为对照)。结果表明:授予柑橘异源四倍体杂种花粉的果实种子败育十分明显,果实瘪籽率达41.4%～96.0%,与对照间的差异极显著(P0.01);但果实单果重、果肉重、果皮重、果皮厚和果形指数与对照间无显著性差异(P0.05);果实可溶性固形物、全糖、Vc和可滴定酸含量与对照也无明显变化,且较适合于瘪籽沙田柚果实生产的体细胞杂种是HR和SR。     

Cytosolic RNA:DNA hybrids activate the cGAS–STING axis

Intracellular recognition of non‐self and also self‐nucleic acids can result in the initiation of potent pro‐inflammatory and antiviral cytokine responses. Most recently, cGAS was shown to be critical for the recognition of cytoplasmic dsDNA. Binding of dsDNA to cGAS results in the synthesis of cGAMP(2′–5′), which then binds to the endoplasmic reticulum resident protein STING. This initiates a signaling cascade that triggers the induction of an antiviral immune response. While most studies on intracellular nucleic acids have focused on dsRNA or dsDNA, it has remained unexplored whether cytosolic RNA:DNA hybrids are also sensed by the innate immune system. Studying synthetic RNA:DNA hybrids, we indeed observed a strong type I interferon response upon cytosolic delivery of this class of molecule. Studies in THP‐1 knockout cells revealed that the recognition of RNA:DNA hybrids is completely attributable to the cGAS–STING pathway. Moreover, in vitro studies showed that recombinant cGAS produced cGAMP upon RNA:DNA hybrid recognition. Altogether, our results introduce RNA:DNA hybrids as a novel class of intracellular PAMP molecules and describe an alternative cGAS ligand next to dsDNA.     

特早熟甘蓝型油菜异交率的测定

利用SSR分子标记技术构建3个特早熟甘蓝型油菜恢复系的指纹图谱,筛选出这3个恢复系的共显性SSR标记,测定了这3个恢复系的异交率。结果表明,恢复系材料4395、3509、4152的异交率分别为46.02%、33.32%、18.12%,在P<0.01时呈极显著差异,说明这3个恢复系的异交差异明显,为确定这3个恢复系在综合杂交种中的比例提供依据。     

Somatic hybridization of sexually incompatible top-fruit tree rootstocks,wild pear (Pyrus communis var. pyraster L.) and Colt cherry (Prunus avium x pseudocerasus)

Summary Mesophyll protoplasts of wild pear (Pyrus communis var. pyraster L., Pomoideae) were chemically fused with cell suspension protoplasts of cherry rootstock Colt (Prunus avium x pseudocerasus, Prunoideae), following an electroporation treatment of the separate parental protoplast systems. Fusion-treated protoplasts were cultured, on modified K8P medium, where it had been previously established that neither parental protoplasts were capable of division. Somatic hybrid calli were recovered and, following caulogenesis on MS medium with zeatin and after rooting of regenerated shoots, complete trees were obtained and grown in vivo. Hybridity of these trees was confirmed based on morphological characters, chromosome complement and isozyme analysis. Two separate cloned lines of this intersubfamilial rootstock somatic hybrid (wild pear (+) Colt cherry) were produced. This is the first report of the production of somatic hybrid plants of two woody species, of agronomic value, within the order Rosales.     

基于孢子形态和分子证据探讨鳞盖蕨属(碗蕨科)系统分类

孢粉学是解决植物分类中疑难类群物种微形态分化的重要方法, 随着分子系统学的发展, 结合这两门学科的优势可以更加有效地解决疑难类群的分类学问题。鳞盖蕨属(Microlepia)是一个分类困难的疑难类群, 采用孢粉学与分子系统学一一对应的方法, 以及居群取样方式, 选取280份样本, 联合4个叶绿体片段(rbcL、trnL-F、psbA-trnH和rps4), 采用最大似然法和贝叶斯法构建该属的系统发生关系, 在此基础上对凭证标本中100份材料的孢子进行观察和分析。综合分子系统学和孢粉学的研究结果, 得出结论: (1) 在形态学研究中广泛被接受的15个物种得到了单系支持, 并厘清了分类困难的复合群; (2) 发现边缘鳞盖蕨(M. marginata)可能存在隐性种; (3) 建议恢复过去归并处理为异名的瑶山鳞盖蕨(M. yaoshanica)、罗浮鳞盖蕨(M. lofoushanensis)、四川鳞盖蕨(M. szechuanica)以及滇西鳞盖蕨(M. subspeluncae); (4) 提出鳞盖蕨属可能存在杂交现象; (5) 提出鳞盖蕨属完整的属下分类建议。     

Transfer of Hessian fly resistance from rye to wheat via radiation-induced terminal and intercalary chromosomal translocations

Summary A new Hessian fly (Mayetiola destructor) resistance gene derived from Balbo rye and its transfer to hexaploid wheat via radiation-induced terminal and intercalary chromosomal translocations are described. Crosses between resistant Balbo rye and susceptible Suwon 92 wheat and between the F₁ amphidiploids and susceptible TAM 106 and Amigo wheats produced resistant BC₂F₃ lines that were identified by C-banding analysis as being 6RL telocentric addition lines. Comparative chromosomal analyses and resistance tests revealed that the resistance gene is located on the 6RL telocentric chromosome. X-irradiated pollen of 6RL addition plants was used to fertilize plants of susceptible wheats TAM 106, TAM 101, and Vona. After several generations of selection for resistance, new sublines were obtained that were homogeneous for resistance. Thirteen of these lines were analyzed by C-banding, and three different wheat-6RL chromosomal translocations (T) were identified. Wheat chromosomes involved in the translocations were 6B, 4B, and 4A. Almost the complete 6RL arm is present in T6BS · 6BL-6RL. Only the distal half of 6RL is present in T4BS · 4BL-6RL, which locates the resistance gene in the distal half of 6RL. Only a very small segment (ca 1.0 m) of the distal region of 6RL is present in an intercalary translocation (Ti) Ti4AS · 4AL-6RL-4AL. The 6RL segment is inserted in the intercalary region between the centromere of chromosome 4A and the large proximal C-band of 4AL. The break-points of the translocations are outside the region of the centromere, indicating that they were induced by the X-ray treatment. All three translocations are cytologically stable and can be used directly in wheat breeding programs.Cooperative investigations of the Kansas Agricultural Experiment Station, Departments of Entomology and Plant Pathology, the Wheat Genetics Resource Center, Kansas State University, and the US Department of Agriculture, Agricultural Research Service. Contribution No. 91-117-JDeceased     

陕油8号种子纯度的RAPD鉴定研究

从杂交油菜“陕油8号”及其亲本中提取基因组DNA,用100个RAPD随机引物进行扩增,从中筛选出3个可将亲本和子代区分的引物BA208、BA1090、BA497。BA208产生亲本互补的特征带BA208-1050bp、BA2081250bp;BA1090产生母本特征带BA1090-700bp,BA497产生父本特征带BA497-870bp,上述谱带均在子代中出现。以BA208产生的特征谱带作为分子标记对杂交油菜种子纯度鉴定得到了一致的结果,并与大田纯度检测结果一致。BA497可将“陕油8号”与当地4个主栽品种有效区分。此外,还对双引物共同鉴定杂交种子纯度问题进行了初步探讨。     

Irregular meiosis in a somatic hybrid between S. bulbocastanum and S. tuberosum detected by species-specific PCR markers and cytological analysis

A system of randomly amplified polymorphic DNA (RAPD) markers was developed to facilitate the transfer of S. bulbocastanum (blb) genes into the S. tuberosum (tbr) genome by hybridization and backcrossing. DNA from tbr, blb and the hexaploid hybrid was used as a template for polymerase chain reaction (PCR) amplification. Polymorphic RAPD products, originating from 10-mer primers, specific for blb were cloned and sequenced at their ends to allow the synthesis of 18-mer primers. The 18-mer primers allowed a more reproducible assay than the corresponding RAPDs. Of eight 18-mer primer pairs, four amplified the expected products specific for blb. However, the stringency of the primer annealing conditions needed to be carefully optimized to avoid amplification of the homeologous tbr product, suggesting that the original RAPD polymorphisms were due to single base-pair changes rather than deletions or insertions. Two primers used for amplification of backcross 2 progeny segregated in a 11 (presence:absence) ratio; the other two were unexpectedly absent. The most likely explanation for the loss of these markers is irregular meiosis in the original hexaploid hybrid and subsequent elimination of chromosomes. Cytological analysis of the meiosis in the hybrid demonstrated widespread irregular pairing and the presence of lagging univalents. In addition, the first backcross individual used as the parent for the second backcross had 54 chromosomes instead of the predicted 60. In conclusion, our results demonstrate that PCR technology can be used for the efficient isolation of taxon-specific markers in Solanum. Furthermore, by the use of these markers we detected the loss of chromosomes that was subsequently shown by cytological analysis to be caused by irregular meiosis of the somatic hybrid.     

斑鳢、乌鳢及其杂种细胞核DNA流式含量分析

以斑鳢(Channa maculata)、乌鳢(C.argus)及其正交杂种斑乌鳢(斑鳢♀×乌鳢♂)和反交杂种乌斑鳢(乌鳢♀×斑鳢♂)的红细胞为材料,以鸡(Gallus gallus)血细胞为DNA标准(2.5 pg/2c,2c指2倍体),采用流式细胞仪测定了这4种鱼的细胞核DNA含量。斑鳢、乌鳢、斑乌鳢及乌斑鳢这4种鱼血细胞DNA的绝对含量分别为(1.488±0.035)pg/2c、(1.489±0.034)pg/2c、(1.522±0.077)pg/2c和(1.520±0.033)pg/2c。斑鳢和乌鳢的细胞核DNA含量差异不显著(P0.05),斑鳢和乌鳢与两种杂交鳢的DNA含量差异显著(P0.05),两种杂交鳢之间的细胞核DNA含量差异不显著(P0.05)。杂交鳢细胞核DNA含量显著高于亲本,可以作为杂种鉴定的依据。     

受精卵反义寡核苷酸显微注射影响因素的研究

以金鱼受精卵为材料,通过胚胎培养探讨反义寡核苷酸显微注射剂量、受精卵质量、胰酶消化浓度等对受精卵反义寡核苷酸显微注射的影响．研究表明,反义寡核苷酸注射剂量在4．6nL、胰酶消化浓度在0.4％的情况下,选用质量较好的受精卵注射,显微注射效果较好,可为之后的基因功能研究提供材料保障．利用显微注射法导入反义寡核苷酸来研究基因的功能．是研究基因功能的有效途径．