Development of two multiplex PCR assays targeting improvement of bread-making and noodle qualities in common wheat

Wheat quality properties are genetically determined by the compositions of high and low molecular weight glutenin subunits, grain hardness, polyphenol oxidase (PPO) activity and starch viscosity. Two multiplex PCR assays were developed and validated using 70 cultivars and advanced lines from Chinese autumn‐sown wheat regions. Multiplex PCR I includes molecular markers for genes/loci ω‐secalin, Glu‐B1‐2a (By8), Glu‐D1‐1d (Dx5), Glu‐A3d, Glu‐B3 (for non‐1B·1R type) and Pinb‐D1b targeting improved gluten parameters and pan bread quality. Multiplex PCR II comprises markers for genes/loci Ppo‐A1, Ppo‐D1 and Wx‐B1b targeting improved noodle quality. The results were consistent with those achieved by SDS‐PAGE and RP‐HPLC, indicating that the two multiplex assays were highly effective, with good repeatability and low costs enabling their use in wheat breeding programmes. In total, nine alleles (subunits) at locus Glu‐B1, four at Glu‐D1 and five at Glu‐A3 locus were identified, and the alleles (subunits) Glu‐B1b (7 + 8), Glu‐B1c (7 + 9), Glu‐D1a (2 + 12), Glu‐D1d (5 + 10), Glu‐A3a, Glu‐A3c and Glu‐A3d were most frequently present in the cultivars and lines tested. The 1B·1R translocation was present in 28 (40.0%) lines, whereas the Wx‐B1 null allele for better noodle quality was present in only seven (10.0%) cultivars and advanced lines, and 37 (52.9%) lines had Pinb‐D1b associated with hard grains. The allele Ppo‐A1b on chromosome 2AL associated with lower PPO activity was present in 38 (54.3%) genotypes, whereas the less effective allele Ppo‐D1a on chromosome 2DL, also associated with low PPO activity was present in 45 (64.3%) of genotypes. These two multiplex PCR assays should be effective in marker assisted selection targeting improved pan bread‐making and noodle qualities.     

N-(1-萘乙酰基)-N'-(4-氨替吡啉基)硫脲(NAT)对小麦生理活性影响的初步研究

以新合成的NAT配制成不同浓度(0.1、1.0、10.50mg·L~(-1))的水溶液,对小麦进行漫种、水培幼苗和大田灌浆期叶面喷洒处理,以与NAT相同浓度的NAA作对比,研究了喷后一定时期小麦叶片生理活性和收割时籽粒千粒重。结果表明,NAT使小麦在不同生长发育时期叶片中的叶绿素含量均增加;幼苗叶片呼吸速率下降;幼苗期低浓度处理NR活性增高,高浓度显著抑制,灌浆期则随处理浓度的升高而显著增加;籽粒千粒重明显高于对照。不论对叶片生理活性还是对籽粒千粒重的影响,NAT的总体效果均明显优于NAA,其中以1 mg·L~(-1)左右浓度的NAT处理效果最佳。     

低温弱光对番茄叶绿素含量变化的影响

研究了低温弱光对中蔬4号和中杂9号番茄苗期植株功能叶叶绿素a、叶绿素b含量和叶绿素a／b变化的影响。结果表明：光照强度为5000 1x或2000 1x时,温度处理为20℃(日)／10℃(夜)、20℃(日)／5℃(夜)、15℃(日)／10℃(夜)和15℃(日)／5℃(夜)对中蔬4号和中杂9号叶绿素a、b含量和a／b值影响较显著,处理时间不同其影响也不同。     

Pre-maturity α-amylase and incipient sprouting in UK winter wheat,with special reference to the variety Rialto

In 1998 and 1999 the UK winter wheat variety Rialto produced unexpected low Hagberg falling numbers that could not be directly linked to sprouting. It was proposed that these reductions in quality could be due to pre-maturity α-amylase activity (PMAA). The problem was not identified during the selection and commercial development stages. Our study tested the hypothesis that the variety Rialto is PMAA-susceptible. Analysis was done on 13 year-location combinations of field grown Rialto. Together, visual and chemical assessments of sprouting and iso-electric focusing of α-amylase isozymes identified several samples with significant α-amylase activity in the apparent absence of sprouting. In addition, tests with α-amylase sensitive Phadebas gel revealed distinctive PMAA discoloration patterns in 10–44% of the grain from the 13 samples, leading to the conclusion that Rialto is PMAA-susceptible. Diurnal temperature range accumulated for an 11 day period during a warm spell in early simulated grain development displayed a significant but negative correlation with the number of grains showing clear PMAA discoloration patterns on Phadebas gel. The number of clear PMAA grains correlated positively with rainfall accumulated over 11 days. These results suggest that PMAA can increase under conditions similar to those conducive to pre-harvest sprouting. It is however also possible that in some instances both PMAA and incipient sprouting could have produced similar patterns of α-amylase activity. In addition to tests with Rialto, Phadebas gel tests were therefore also done with the known high Hagberg varieties Option and Malacca, sprouted in a controlled environment. Results from the additional gel tests suggest that visual and chemical assessments of sprouting in the grain combined with Phadebas gel analysis could identify PMAA more reliably in grain sub-samples than Phadebas gel analysis alone.     

超高产两系杂交稻不同施氨水平剑叶光合特性研究

研究了超高产两系杂交稻两优培九、金两优36在不同施氮水平下剑叶的光合特性，结果表明，两优培九、金两优36在每公顷施氮120kg、165kg、210kg 3种水平下，剑叶比叶重、叶绿素含量、光合速率均比对照汕优63高，这可能是两优培九、金两优36高产的生理基础之一。在每公顷施氮120-210kg的范围内，两优培九剑叶光合速率及产量均随施氮水平的增加而提高，金两优36剑叶光合速率及产量在公顷施氮165kg最高，增加到210kg时反而下降。     

光质对番茄果实表面光系统活性的影响

以保罗塔番茄果实为试材,采用叶绿素成像荧光仪(MINI-IMAGING-PAM)测试分析了蓝(453±2)nm、绿(519±2)nm、红(623±2)nm 3种光质对番茄果实表面光化学活性的影响。结果表明:随着光强的增加(200～2 500μmol/(m.2s)),蓝光和红光处理番茄果实表面光系统Ⅱ(PSII)最大光化学量子产量Fv/Fm、PSII实际量子产量Y(Ⅱ)、电子传递速率ETR和PSII天线转化效率Fv’/Fm’均呈"S"型下降趋势;光系统间激发能分配不平衡偏离系数β/α-1急剧上升,反映出高光强导致光系统间激发能分配的不平衡,PSⅡ和PSⅠ间线性电子传递的协调性降低;而绿光处理番茄果实表面叶绿素荧光参数相对来说变化不大,可能是绿光减少了叶绿素对光能的过度吸收,使得光抑制程度较轻;蓝光处理番茄果实表面Fv/Fm、Y(Ⅱ)、ETR和Fv’/Fm’均小于红光和绿光处理,并且随着光强的增加这种差距越来越明显,表明蓝光对光抑制更敏感,更容易受到强光胁迫的影响;另外,当光强为200～1 000μmol/(m.2s)时,番茄果实表面Fv/Fm、Y(Ⅱ)和Fv’/Fm’值红光>绿光,超过1 000μmol/(m.2s)时,则绿光>红光,反映出绿光处理番茄果实适应强光的能力较强。     

不同株穗型水稻超高产品种叶绿素含量变化规律及籽粒灌浆动态研究

对不同株、穗型水稻超高产品种叶绿素含量变化规律及籽粒灌浆动态研究结果表明：（1）株、穗型较好的半直立穗型品种上、中、下三部籽粒收获,籽粒重量相差无几,分布均匀,灌浆速率高,灌浆物质充足,弱势粒得到了较好的充实;直立穗型品种,中部籽粒重量较小,弱势粒灌浆不足;大散穗型超高产品种为明显的异步灌浆,上、中     

风箱果叶色变化及呈色机理研究

为研究风箱果叶片不同生长发育过程中叶色变化及呈色机理的问题,以风箱果、‘金叶’风箱果、‘紫叶’风箱果为材料,对其叶片从幼嫩到成熟转变过程5 个生长阶段的叶部特征和叶色变化进行比较研究,并采用徒手切片法观察叶片中色素空间分布。结果表明,随着3 种风箱果的生长,叶片面积均呈逐渐增大的趋势,‘紫叶’风箱果叶面积最大;3 种风箱果叶绿素a 含量、叶绿素b 含量、叶绿素/类胡萝卜素均呈现逐渐升高的趋势,风箱果不同生长阶段叶绿素a 含量、叶绿素b 含量均显著高于‘紫叶’风箱果和‘金叶’风箱果;3 种风箱果叶片色素均主要分布于栅栏组织内,在海绵组织和表皮细胞分布较少。上述结果明确了风箱果叶片在生长发育过程中的叶部特征和叶色表现,为其科学合理应用提供了理论依据。     

麦玉两熟秸秆长期全量还田模式下氮肥对冬小麦生长发育、产量及品质的影响

为了探讨小麦-玉米两熟区长期秸秆还田模式下氮肥的适宜用量,采取秸秆不还田、秸秆全量还田为主因素,氮肥用量（设0、90、180、270、360 kg/hm2 5 个水平）为辅因素的裂区试验设计,研究了麦玉两熟秸秆长期还田配施氮肥对冬小麦生长发育、产量及品质的影响。研究结果表明：同一氮肥水平秸秆还田处理能够增大叶面积,增强光合作用,提高叶绿素含量及地上干物质量,增加产量。随施氮量的增加,叶绿素呈增长趋势,但叶面积、地上干物质量、穗数、穗粒数及产量均呈先增后减的趋势。蛋白质及湿面筋含量随施氮量增加而提高。过量施用氮肥(270、360 kg/hm2),产生盐害,会抑制小麦生长,导致减产。秸秆还田配施180 kg/hm2氮肥处理能明显促进冬小麦的生长发育,提高产量和品质。经模拟分析,秸秆全量还田模式下小麦最高产量的推荐施氮肥量为216.11 kg/hm2。     

PRRSV 2b蛋白与LGALS1相互作用的验证研究

旨在研究猪繁殖与呼吸综合症病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)2b蛋白与宿主细胞蛋白之间的相互作用。本研究利用酵母回返试验、GST-pull down试验、免疫共沉淀试验3种方法对LGALS1与PRRSV 2b蛋白间是否相互作用进行验证。结果表明:在酵母回返试验中,含有LGALS1与2b基因的杂交酵母菌株在SD/-Trp-Leu-His-Ade/AbA/X-α-gal的平板上生长出蓝色菌落;在GST-pull down试验中,LGALS1与2b蛋白反应的样品经Western blot可检测到融合蛋白带;在免疫共沉淀试验中,LGALS1与2b蛋白反应的样品经HA单抗沉淀后,经Western blot可检测到相应蛋白带。该研究证明了LGALS1与PRRSV 2b蛋白间发生相互作用,为研究PRRSV感染途径和增殖过程提供了理论基础。