Intra-specific variation of wheat grain quality in response to elevated [CO2] at two sowing times under rain-fed and irrigation treatments

In order to investigate the intra-specific variation of wheat grain quality response to elevated atmospheric CO₂ concentration (e[CO₂]), eight wheat (Triticum aestivum L.)cultivars were grown at two CO₂ concentrations ([CO₂]) (current atmospheric, 389 CO₂ μmol mol⁻¹vs. e[CO₂], FACE (Free-Air CO₂ Enrichment), 550  ±  10% CO₂ μmol mol⁻¹), at two water levels (rain-fed vs. irrigated) and at two times of sowing (TOS₁, vs. TOS₂). The TOS treatment was mainly imposed to understand whether e[CO₂] could modify the effects of timing of higher grain filling temperatures on grain quality. When plants were grown at TOS₁, TKW (thousand kernel weight), grain test weight, hardness index, P, Ca, Na and phytate were not significantly changed under e[CO₂]. On the other hand, e[CO₂] increased TKW (16%), hardness index (9%), kernel diameter (6%), test weight (2%) but decreased grain protein (10%) and grain phytate (11%) at TOS₂. In regard to grain Zn, Mn and Cu concentrations and some flour rheological properties, cultivar specific responses to e[CO₂] were observed at both sowing times. Observed genetic variability in response to e[CO₂] in terms of grain minerals and flour rheological properties could be easily incorporated into future wheat breeding programs to enable adaptation to climate change.     

Quantitative proteomic analysis of wheat grain proteins reveals differential effects of silencing of omega-5 gliadin genes in transgenic lines

Novel wheat lines with altered flour compositions can be used to decipher the roles of specific gluten proteins in flour quality. Grain proteins from transgenic wheat lines in which genes encoding the omega-5 gliadins were silenced by RNA interference (RNAi) were analyzed by quantitative 2-dimensional gel electrophoresis (2-DE). The precise effects of the genetic modifications on the proteome were assessed in four homozygous lines generated with the same RNAi construct. Two of the lines showed >80% decreases in omega-5 gliadins with only small changes in the levels of other gluten proteins. In the other two lines, omega-5 gliadins were not detectable by 2-DE. However, there were notable reductions in all omega-1,2 gliadins. Small decreases in several other gluten proteins were also detected in one of the lines. The other line showed notable decreases in three HMW-GS and an s-type LMW-GS, increases in two m-type LMW-GS and several alpha gliadins, as well as increases in both serpins and triticin. The study demonstrates that the same RNAi construct can have differential effects on the wheat grain proteome and highlights the importance of detailed proteomic analyses of transgenic grain prior to selecting lines for further assessment of flour quality and allergenic potential.     

温敏雄性不育小麦BNS366花粉败育的细胞学观察

为了明确温敏雄性不育小麦BNS366雄性败育的细胞学机理,采用醋酸洋红染色法观察减数分裂和小孢子发育过程,用1%I2-KI溶液染色进行花粉育性统计;以温敏不育系BNS和常规品种扬麦13作为对照。结果表明,BNS366的减数分裂和小孢子发育过程与BNS完全一致,即:在减数分裂过程中,中期Ⅰ染色体出现滞后现象,在形成二分体和四分体时细胞质分裂不均匀,出现三孢现象;小孢子发育过程中,只观察到单核靠边期,最后核物质解体出现空孢现象。而扬麦13在减数分裂过程中染色体表现正常,小孢子发育过程中,观察到单核期→双核期→三核期。开花期,BNS366和BNS花粉均表现典败和圆败,扬麦13可育。减数分裂表现出的异常行为是导致BNS366花粉败育的原因之一,单核靠边期是其败育的关键时期。     

不同种类真菌多糖对小麦黄矮病的防治效果

为了筛选小麦黄矮病的真菌多糖防治药剂及其适宜浓度,将云芝、灵芝、虫草等13种真菌多糖稀释成0.1、0.5、1.0、2.0g·L-1溶液,以清水为对照,比较分析了叶面喷施真菌多糖对小麦黄矮病的预防和治疗效果,同时对防效好的1.1%云芝葡聚糖(多糖)与常规植物抗病毒药剂的田间防效进行了比较。结果表明,13种真菌多糖对小麦黄矮病的预防作用都显著高于治疗作用,且真菌多糖间和浓度间差异均显著,其中云芝多糖1.0g·L-1、云芝多糖2.0g·L-1、蜜环菌多糖2.0g·L-1、灰树花多糖2.0g·L-1、猪苓多糖2.0g·L-1的预防效果分别为86.88%、86.58%、86.33%、86.04%和85.97%。1.1%云芝葡聚糖800倍液为适宜喷施浓度,其防效64.27%,保产22.70%,说明云芝多糖是一种有前途的新型小麦黄矮病免疫抑制剂。     

胶州市小麦禾谷孢囊线虫群体动态的年度间差异

为明确山东胶州市禾谷孢囊线虫(Heterodera avenae,CCN)的田间发生状况及其年度间的差异,于2011-2012和2012-2013年两个小麦生长季在该地区麦田进行定点定期取样、土壤中线虫分离和根组织内线虫染色后镜检,以确定各虫态的季节和年度间群体动态变化。结果表明,与2011-2012年相比,2012-2013年春季CCN 2龄幼虫(J2)孵出较早,但对根系侵入较晚;土壤中J2群体密度较高,根组织中反而较低;根内幼虫发育不良,表现为3龄和4龄幼虫发生较晚且持续时间较短,没有明显发生高峰,群体密度持续低下;小麦收获后土壤中孢囊密度减少85.9%。2012-2013年J2侵入期推后和侵入率下降与3-4月份土壤干旱有关,而根内幼虫发育不良与4-5月份气温偏低有关。因此CCN侵染和发育关键期的干旱和低温可能是造成孢囊密度下降的主要原因。     

不同小麦品系耐低钾性的综合评价

为了鉴定并筛选耐低钾的小麦品系,以小麦(花培3号×豫麦57)双单倍体(DH)群体(168个株系)为材料,测定低钾处理和对照(正常钾处理)下,不同株系的地上部干重、地下部干重、整株干重、根冠比、根平均直径、根体积、根总表面积、总根长、根系活力、SOD活性、POD活性、钾浓度和钾累积量,分析其在低钾胁迫下的反应,综合评价不同品系的耐低钾性,筛选可用于评价耐低钾性的指标。结果表明,单一指标不能全面、准确地反映小麦的耐低钾性;通过主成分分析和隶属函数分析相结合的方法,获得各供试材料耐低钾性的综合评价值(D值),且D值可以直接反映各DH株系耐低钾能力的大小;通过逐步回归的方法建立了最优回归方程,筛选出6个与小麦耐低钾性显著相关的指标。     

小麦-黑麦小片段易位的快中子辐照诱导

为研究快中子辐照对新合成的小麦-黑麦双二倍体染色体结构变化的影响,分别利用5Gy、15Gy和20Gy三种快中子辐照剂量对其种子进行辐射诱变,采用基因组原位杂交(GISH)技术对M0和M1种子的根尖细胞进行了检测。结果表明,三种剂量下发生小麦-黑麦染色体易位的总频率为12.28%,其中发生外源小片段易位、外源大片段易位和罗伯逊易位的频率分别为9.36%,5.85%和2.34%。三种剂量都可引起小麦与黑麦染色体之间发生易位,分别有1.28%、9.04%和44.94%的M1种子产生了易位。说明随着快中子辐照剂量的增加,小麦与黑麦染色体间发生易位的频率也在增高。本研究为外源染色体片段(或优异基因)导入小麦品种提供了一种高效的技术手段。     

低温下硅对春小麦种子萌发及抗氧化和渗透调节系统的影响

为了解硅缓解低温胁迫春小麦种子萌发的作用,以春小麦品种龙麦26和克旱16为材料,分析了低温条件下硅酸钠处理后春小麦种子萌发、渗透调节物质含量和保护酶活性的变化。结果表明,低温条件下硅酸钠处理后,春小麦种子萌发显著受到促进,发芽率、发芽势、发芽指数和活力指数提高,其中以1.0mmol·L-1硅酸钠处理效果最好;小麦种子α-淀粉酶、超氧化物歧化酶、过氧化物酶活性以及可溶性糖含量显著提高,丙二醛含量、外渗电导率和脯氨酸含量显著降低。说明外源硅可缓解低温对春小麦种子萌发产生的影响。     

小麦籽粒淀粉含量的QTL定位及效应分析

为深入了解小麦籽粒淀粉含量的遗传特性和QTL效应,选择淀粉含量差异较大的小麦品种M344和武春3号杂交,创制F2∶3群体,并利用973对SSR引物对小麦籽粒总淀粉、直链淀粉和抗性淀粉含量进行基于混合线性模型的QTL定位及效应分析。构建了包含有163个标记的遗传连锁图谱,分布于18条染色体,连锁图谱总长度为1 622.5cM,标记间平均距离为9.95cM。QTL分析表明,共检测到4个主效QTL和12对上位性QTL,涉及1B、2B、2D、4A、5D、6A和7A等16条染色体。其中,抗性淀粉含量上位性QTL3对,总淀粉含量上位性QTL 4对,直链淀粉含量上位性QTL 5对,总贡献率分别为8.34%、17.50%和8.13%。总淀粉和直链淀粉含量各检测到1个加性QTL,抗性淀粉含量检测到2个加性QTL,贡献率分别为5.34%、8.49%、11.47%、12.53%。研究发现,2B染色体Xwmc453.3~Xcfd44.2标记区间的QTL位点同时影响抗性淀粉和总淀粉含量,2D染色体Xgwm296~Xgwm455.2标记区间的QTL位点同时影响抗性淀粉和直链淀粉含量,7A染色体Xwmc488.1~Xwmc488.2标记区间的QTL位点同时影响直链淀粉和总淀粉含量。     

小麦PDS基因VIGS载体构建体系的优化及验证

为在小麦上进行VIGS体系的优化,根据GenBank中公布的小麦PDS基因序列(FJ517553.1)设计特异引物,利用RT-PCR技术克隆PDS基因片段,以BSMV:γ-2a为原载体,通过酶切将原载体中的2a基因片段替换为PDS基因片段,构建小麦PDS基因的VIGS重组载体BSMV:γ-PDS。经测序确认,其与GeneBank中登录的小麦PDS基因序列同源性为97%。重组载体经本实验室简化改良的方法进行转录模板的处理。与试剂盒所提供的方法相比,本研究将RNA酶孵育处理步骤整合入质粒提取的步骤中,省略了SDS-蛋白酶K孵育处理等步骤,降低了引入新杂质的风险,同时降低了实验成本,节省了时间和精力。体外转录的电泳结果表明,体外转录反应的产物浓度大,条带单一,可用于接种实验。抽穗期小麦穗部接种实验及实时定量PCR的结果均表明,小麦内源PDS基因被有效沉默,证实了优化的重组载体BSMV:γ-PDS体系的有效性,奠定了利用VIGS载体侵染抽穗期小麦体系优化的基础。