Development of a kompetitive allele‐specific PCR marker for selection of the mutated Wx‐D1d allele in wheat breeding

Waxy (Wx) protein is a key enzyme for synthesis of amylose in endosperm. Amylose content in wheat grain influences the quality of end‐use products. Seven alleles have been described at the Wx‐D1 locus, but only two of them (Wx‐D1b, Wx‐D1e) were genotyped with codominant markers. The waxy wheat line K107Wx1 developed by treating ‘Kanto 107’ seeds with ethyl methanesulphonate carries the Wx‐D1d allele. However, no molecular basis supports this nomenclature. In the present study, DNA sequence analysis confirmed that a single nucleotide polymorphism in the sixth exon of Wx‐D1 changed tryptophan at position 301 into a termination codon. Based on this sequence variation, a PCR‐based KASP marker was developed to detect this point mutation using 68 BC₈F₁ plants and 297 BC₈F₂ lines derived from the cross ‘Ningmai 14’*9/K107Wx1. Combined with codominant markers for the Wx‐A1 and Wx‐B1 alleles, waxy and non‐waxy near‐isogenic lines were distinguished. The KASP marker was efficient in identifying the mutant allele and can be used to transfer waxiness to elite lines.     

葡萄‘小白玫瑰’及其突变品种‘小红玫瑰’果皮颜色变异机理分析

‘小红玫瑰’葡萄是源自于‘小白玫瑰’的突变品种,二者都是优良的酿酒葡萄品种,但其果实颜色变异的分子机理尚不清楚。用12对SSR(Simple sequence repeats)荧光标记特征引物对两品种进行分析,结果表明这些位点在二者之间无差别。对色泽变异决定基因Vvmyb A1的基因型进行分析,‘小白玫瑰’仅检测到含有插入逆转座子Gret1(grapevine retrotransposon 1)的VvmybA1a,说明其为VvmybA1a/VvmybA1a纯合体;而‘小红玫瑰’检测到了Vvmyb A1a和残留Gret1 3′-LTR(long terminal repeat)的VvmybA1b,说明其为VvmybA1a/VvmybA1b的杂合体。花色苷合成相关基因的表达分析表明,VvmybA1、UFGT、F3′5′H、CHS、GST和OMT在‘小红玫瑰’及有色对照品种‘黑比诺’中大量表达,而在‘小白玫瑰’和无色对照品种‘白比诺’中几乎不表达。通过HPLC–ESI–MS/MS对花色苷主要成分进行测定,结果表明矢车菊素3–O–葡萄糖苷(CyG)、矢车菊素3,5–O–双葡萄糖苷(Cy2G)、飞燕草素3–O–葡萄糖苷(DpG)、锦葵色素3–O–葡萄糖苷(MvG)、芍药色素3–O–葡萄糖苷(PnG)和矮牵牛色素3–O–葡萄糖苷(PtG)等6种花色苷在有色品种‘小红玫瑰’和‘黑比诺’果皮中的含量均显著高于无色品种,花葵素3–O–葡萄糖苷(Pg G)、花葵素3,5–O–双葡萄糖苷(Pg2G)在有色/无色品种间无显著差异,且含量很低。说明‘小红玫瑰’的着色是由于转录因子基因Vvmyb A1及其调控结构基因的表达引起的,而无功能的VvmybA1a等位基因突变为有功能的VvmybA1b等位基因是其果皮颜色变异的遗传学原因。     

竹炭固定化紫外诱变假单胞菌处理间甲酚废水的研究

用紫外光对假单胞菌株进行诱变,以竹炭为载体,将紫外诱变假单胞菌固定在竹炭上,用竹炭固定化紫外诱变假单胞菌处理间甲酚水样。考察竹炭固定化紫外诱变假单胞菌投加量和水样pH值对间甲酚去除的影响以及进水浓度随反应时间的变化关系,研究竹炭固定化紫外诱变假单胞菌去除间甲酚的反应动力学。结果表明:相对于原菌株,菌株经紫外诱变后,生长周期缩短了6h。经紫外照射120s的假单胞菌可以在竹炭表面及内部孔隙形成明显菌胶团,诱变菌在竹炭上所成的生物量明显较未经诱变菌增加。竹炭固定化紫外诱变假单胞菌能有效地去除水样中间甲酚。竹炭固定化紫外诱变假单胞菌投加量和水样pH值影响到间甲酚的去除效果,pH值在4～6时,间甲酚的去除效果较好。20g竹炭固定化紫外诱变假单胞菌处理100mL初始浓度50,100,120,150,180mg·L-1间甲酚模拟水样42h,去除率依次为90.9%,76.4%,72.9%,64.6%和49.7%。竹炭固定化紫外诱变假单胞菌对间甲酚的去除能较好地符合零级反应方程。     

大豆化学诱变育种及其规律的研究

应用化学诱变剂处理大豆种子,对M_1代植株的生育及形态均有强烈的抑制作用.M_1代所产生的形态变异主要是由于化学物质扰乱了植株生理而产生生理损伤所致.在M_2代植株各性状中以分枝数、株高、单株荚数的遗传变异系数较大,说明这些性状选择的潜力是大的,通过选择容易达到预期目标.诱变M_3代绝大多数性状仍在分离.加强选择,能选出早熟、大粒、农艺性状好、丰产的新品系.     

Parial resistance in the barley (Hordeum vulgare L.) cultivar 'Chikurin Ibaraki 1' to two PAV-like isolates of barley yellow-dwarf virus: allelic variability at the Yd2 gene locus

The Japanese barley cultivar, ‘Chikurin Ibaraki 1’, is partially resistant to the PAV serotype of barley yellow-dwarf virus (BYDV), but its induced mutant line, Ea52, is susceptible. The inheritance of resistance in cv. ‘Chikurin Ibaraki 1’ to BYDV-PAV was investigated. The F, and F2 plants of crosses of cvs ‘Chikurin Ibaraki 1’, Ea52, ‘Vixen’, carrying the Yd2 gene of resistance, and ‘Plaisant’, a susceptible French cultivar, were tested in growth chamber and field conditions. Isolate RG, against which ‘Chikurin Ibaraki 1’ is partially resistant in growth chamber and field conditions, and isolate 2t, which overcomes the partial resistance of ‘Chikurin Ibaraki 1’ in field conditions (Chalhoub et al. 1994) were used. The segregation of F2 plants of crosses between ‘Chikurin Ibaraki 1’ and the susceptible cultivars to isolate RG (one resistant to three susceptible) suggests that the resistance of ‘Chikurin Ibaraki 1’ is controlled by a single recessive gene. All 537 F2 plants of ‘Chikurin Ibaraki 1’בVixen’ tested with isolate RG in growth chamber and field conditions were resistant. The F2 plants of this cross were all resistant to isolate 2t in growth chamber conditions but segregated with a ratio of one resistant to three susceptible in field conditions owing to the susceptibility of ‘Chikurin Ibaraki 1’ to this isolate. Results suggest that the resistance gene in ‘Chikurin Ibaraki 1’ is tightly linked or allelic with the Yd2 gene in ‘Vixen’. However, it differs from this gene in ‘Vixen’ in that it can be overcome by isolate 2t in field conditions.     

Mutation genetics of salt tolerance in barley: An assessment of Golden Promise and other semi-dwarf mutants

A review of research at the Scottish Crop Research Institute (SCRI) on the effects of semi-dwarfing genes on salt tolerance in barley is given. Work began in1993 with the fortuitous and unexpected result that the cultivar ‘Golden Promise’ showed considerable tolerance to salt. Golden Promise is a gamma-ray induced semi-dwarf mutant of the cultivar ‘Maythorpe’. The parent and mutant cultivars are presumed to be isogenic, but show significant differences in their responses to salt stress. The positive and pleiotropic effects of the mutant gene, commonly known as GPert were found to be effective in a number of genetic backgrounds. Earlier, in 1991 Frackowiak showed that the GPert mutation was allelic to the ari-e mutants in barley. The ari-emutants were salt tested and found to show the same positive responses to salt stress as Golden Promise. This supported the allelism tests, and consequently the GPert symbol was changed to ari-e.GP. The semi-dwarf mutant sdw1 (also known as denso) and the erectoides semi-dwarf mutant,ert-k ³² were also tested for their effects on tolerance to salt, but did not show any positive effects. Salt tolerance was therefore not a general phenomenon of semi-dwarf stature but specific to mutations at the Ari-e locus in these lines. Genetic markers (RAPDs, AFLPs and SSRs) have been used for fingerprinting, genetic mapping, and QTL analysis. The markers have helped 1) confirm the isogenic relationship between Maythorpe and Golden Promise, 2)clarify the confusion over the pedigree of Golden Promise, and 3) genetically map the ari-e.GPlocus and examine its pleiotropic effects. This revised version was published online in August 2006 with corrections to the Cover Date.     

Increasing mutagenic efficiency of gamma-radiation in rice by irradiation of seeds at extremely low temperature

Summary Dry seeds of rice, Oryza sativa L., containing about 13 percent moisture, were exposed to ⁶⁰Co gamma raysat extremely low temperature (–196°C). M₁ damage on survival rate, seedling height and seed set were found to become much smaller in the cooling treatment than in the not-cooled control. Chlorophyll and heading date mutation frequencies in the M₂ generation were not affected by the decreased M₁ damage caused by the cooling treatment, and the mutation frequencies could be significantly increased by applying higher doses combined with the cooling treatment. Frequency of sterile or semi-sterile mutants in the M₂ generation was found to be significantly lower in the cooling treatment than in the not-cooled control.It is assumed from the experimental results that extremely low temperatures during irradiation could reduce the amount of physiological damage and gross chromosome aberrations. This results in an increased efficiency of producing minute deletions and/or point mutations.     

Prospects of polyploidizing cassava, Manihot esculenta Crantz, by unreduced microspores

Eleven interspecific hybrids of cassava with the wild relatives Manihot glaziovii, M. pseudoglaziovii. M. dichotoma and M. anomala were examined to detect unreduced microspores. The frequency of dyads and triads was counted, as well as pollen viability. Of these hybrids, the second generation progeny of cassava×M. glaziovii showed a high frequency of unreduced microspores (3.7%) permitting their selection for further manipulation of this character. An association of vegetatively-reproduced genotypes and the occurrence of unreduced microspores has also been observed.     

Induction,genetics and possible use of glabrousness in chickpea

Summary A giabrous mutant was identified from progenies of chickpea seeds that were treated with ethyl methane sulphonate (EMS). The mutant has no shoot hairs in contrast to the dense hairs on normal chickpeas. The character is governed by a single recessive gene. This mutant can be useful in certain pathological and eniomological studies.     

Specific developmental stages of gametogenesis for radiosensitivity and mutagenesis in rice

Summary Developmental stages during gametogenesis of rice were histologically examined in the period from differentiation of reproductive organs to anthesis. Plants were exposed to acute X-rays of 20 Gy. Radiosensitivity and mutation frequency were investigated in relation to the developmental stages of reproductive organs. The most radiosensitive stage, as measured by reduction of the M1 pollen-and seed-fertilities, was the last premeiotic interphase. Mutations induced at different developmental stages were scored in M3 strains. Sterility mutants and short-culm mutants were most frequently observed. Grain shape, panicle morphology, heading-date and endosperm character mutants were induced at a relatively low frequency. The overall mutation frequency varied with the developmental stage at the time of irradiation. The highest overall mutation frequency was observed when radiation was applied 10 days before anthesis, the late tetrad stage of microspores. Radiation exposure of florets at the late tetrad stage was found to be a more efficient method of inducing a large number of mutations than radiations applied to seeds and fertilized egg cells.