农杆菌介导大麦无筛选标记转基因植株的获得

【目的】目前,国内外大麦遗传转化主要利用Golden Promise品种,基因依赖性严重,尤其是大麦的转化效率较低,并且获得安全型转基因大麦植株对其进一步产业化非常重要。建立高效、无筛选标记大麦遗传转化体系,拓展大麦遗传转化的受体基因型,为大麦基因功能解析和大麦转基因育种及商业化种植提供技术保障。【方法】以优良大麦品种Vlamingh为受体,取开花授粉后14 d左右的幼胚为转化材料,通过对培养基成分及培养步骤优化,建立农杆菌介导的高效遗传转化体系,并利用该体系将Bar和GUS在不同T-DNA区段的双T-DNA表达载体pWMB123转化大麦,获得候选转基因植株,然后利用PCR、Bar试纸条、组织化学染色和Southern blot等检测方法,在T₁代转基因植株中成功获得无筛选标记大麦转基因植株。【结果】在愈伤组织分化阶段,发现培养基中添加1.0 mg·L^-1 KT、0.5 mg·L^-1 6-BA和0.05 mg·L^-1 NAA明显促进愈伤组织分化。在转基因植株生根阶段,发现采用添加1.0 mg·L^-1的IBA的SM1（无其他生长素）的生根效果最佳,培养基中添加2.5 mg·L^-1 CuSO₄显著降低了大麦转基因植株白化现象。共转化了138个幼胚,最终获得14株大麦转基因植株,转化效率10.14%。PCR、Bar试纸条、GUS染色等检测证实,T₀代转基因植株中均含有Bar,而仅有10株含有GUS,2个T-DNA的共转化效率为71.43%。选取4个同时含有Bar和GUS的转基因植株,对其自交后代进行检测,在BL8株系中筛选到2株只含GUS而不含Bar的转基因植株,无筛选标记效率为6.9%。在T₁代转基因植株中对Bar和GUS进行了Southern blot鉴定,发现在多数转基因植株中Bar和GUS均为多拷贝整合,进一步证实BL8-15和BL8-19为无筛选标记的转基因植株。【结论】利用大麦品种Vlamingh为转化材料可以较高效率获得转基因植株,提高愈伤组织分化效率和转基因植株生根效率,降低转基因植株白化现象。利用农杆菌介导双T-DNA表达载体转化大麦,成功获得了无筛选标记转基因植株。     

施肥对春青稞干物质积累、分配及产量的影响

为研究施肥对青稞干物质积累、分配及产量的调节作用,以‘藏青27’、‘QTB13’和‘QTB25’为试验材料,比较分析不同施肥处理下干物质积累、分配及产量的变化规律。结果表明：增加施肥量促进青稞分蘖期—成熟期的干物质积累及开花期和成熟期干物质向营养器官和籽粒的分配,提高了花前营养器官贮藏同化物转运量及对籽粒贡献率,降低了花后同化物输入籽粒量对籽粒贡献率。‘QTB13’和‘QTB25’在F₂条件下,更有利于干物质积累及向营养器官和穗部的分配,花后同化物输入籽粒量最大,产量也最大。‘藏青27’在F₃条件下,更有利于干物质积累及向营养器官和穗部的分配,花后同化物输入籽粒量和对籽粒贡献率最大,产量也最大。说明合理施肥有利于青稞干物质积累分配及产量提高。     

Transport of cadmium from soil to grain in cereal crops: A review

Due to rapid urbanization and industrialization, many soils for crop production are contaminated by cadmium(Cd), a heavy metal highly toxic to many organisms. Cereal crops such as rice, wheat, maize, and barley are the primary dietary source of Cd for humans, and reducing Cd transfer from soil to their grains is therefore an important issue for food safety. During the last decade, great progress has been made in elucidating the molecular mechanisms of Cd transport, particularly in rice. Inter-and intraspecific variations in Cd accumulation have been observed in cereal crops. Transporters for Cd have been identified in rice and other cereal crops using genotypic differences in Cd accumulation and mutant approaches. These transporters belong to different transporter families and are involved in the uptake, vacuolar sequestration, root-to-shoot translocation, and distribution of Cd. Attempts have been made to reduce Cd accumulation in grains by manipulating these transporters through overexpression or knockout of the transporter genes, as well as through marker-assisted selection breeding based on genotypic differences in Cd accumulation in the grains. In this review, we describe recent progress on molecular mechanisms of Cd accumulation in cereal crops and compare different molecular strategies for minimizing Cd accumulation in grains.     

种植密度对2个青稞品种抗倒伏及秸秆饲用特性的影响

种植密度是影响青稞抗倒伏和秸秆饲用特性的重要因子。以抗倒伏品种昆仑14号和倒伏品种门源亮蓝为试验材料,比较研究种植密度对这2个品种生长发育、抗倒伏特性和秸秆饲用特性的影响。结果表明,种植密度对2个品种的抗倒伏和秸秆饲用特性的影响存在差异。随着种植密度的增加,昆仑14号根长、根体积、根数和根干重先增后降,茎粗和壁厚依次下降;门源亮蓝根系和茎秆相关指标则随种植密度增大而下降。昆仑14号抗倒伏相关指标先增后降,但整个生育期未发生倒伏;门源亮蓝各指标均显著降低,诱发倒伏现象提前发生,致使倒伏率增大、倒伏程度加剧。昆仑14号茎秆中性洗涤纤维、酸性洗涤纤维、半纤维素、纤维素和木质素等化学成分含量随密度增加先增后降,门源亮蓝各成分含量呈下降趋势,相对饲喂价值随密度增加呈现增高的趋势。综合抗倒伏特性与秸秆饲用特性,昆仑14号最佳种植密度为375×104株hm~(-2),门源亮蓝粮饲兼用时适宜密度为300×104～375×104株hm~(-2)。     

播期对麦(油)后直播棉产量、品质及氮磷钾利用的影响

为明确长江流域下游棉区麦(油)后直播棉生产的适宜播期,本研究以?中棉所50‘为材料,于2017—2018年进行大田试验,研究不同播期(5月15日、5月25日、6月4日、6月14日和6月24日)对直播棉产量、品质及养分利用的影响。结果表明,播期推迟,棉花生育进程推迟;花铃期日均温和日照时数降低、有效积温先升高后降低且以6月4日播期花铃期有效积温最高。可见,前3个播期棉花花铃期温光条件较好。棉株与生殖器官的生物量及氮磷钾累积量随播期推迟降低,且前3个播期的棉株生物量和养分累积进入较高速率累积期早于后2个播期的棉株。随着播期的推迟,直播棉成铃数与皮棉产量显著降低。与5月15日播期相比,其余4个播期直播棉的皮棉产量分别降低15.9%～16.9%、13.1%～16.9%、26.9%～33.5%、58.2～62.0%。100kg皮棉氮、磷和钾吸收量随播期推迟而增加,但养分利用效率降低。不同播期间,以6月4日和6月14日播期棉纤维综合品质较优。综上,本试验条件下,5月15日—6月4日是长江流域棉区麦(油)后直播棉获得较高产量的适宜播期,该播期内适当推迟播种有利于棉花优质纤维的形成。     

Phenotypic and allelic variation for wort protein Z in Australian and Canadian barleys

Protein Z is a major component in beer foam. Two-dimensional electrophoresis was used to analyze wort proteins of two Australian (Buloke and Commander) and two Canadian (CDC Meredith and Bentley) varieties. The Canadian barley contained more abundant proteins from MW 40–45 kDa (pI 5 to 7). These proteins were identified as either protein Z4 or protein Z7 using liquid chromatography–mass spectrometry. Full-length gene of protein Z4 and Z7 were sequenced from Canadian and Australian barleys. Sequence differences were identified in the coding region and upstream regions of the two genes, resulting in protein sequence and expression variations. Molecular markers were designed according to the indels in the upstream regions of protein Z4 and Z7 genes. These markers were highly correlated to wort protein Z content in Canadian and Australian varieties. The Canadian barleys contained ‘high level’ genotypes for protein Z4 and Z7 while most Australian barleys had ‘low level’ genotypes for protein Z4, Z7 or both. The markers identified in this study provide a valuable tool for the selection of protein Z alleles in marker-assisted breeding. Total protein Z content was assessed using different steeping conditions, and increasing air-rest time increased protein Z content in 15 varieties.     

西藏地区青稞籽粒营养品质分析及评价

为探明西藏地区青稞品种间的性状差异及性状间的相关性，筛选优异品质青稞种质，以251个西藏青稞种质资源为材料，测定了籽粒淀粉、粗蛋白、β-葡聚糖、γ-氨基丁酸、大量元素（Ca、P）、微量元素（Fe、Zn、Se）含量共9个品质指标，分析了被测指标间关系。结果表明，所测定的9项品质指标在供试种质资源间均存在不同程度的差异，淀粉含量的变异系数最小，Se含量的变异系数最大；粗蛋白与淀粉含量呈极显著负相关（r= -0.54），粗蛋白与γ-氨基丁酸、Ca、P、Zn、Fe含量均呈极显著正相关。对被测指标进行主成分分析发现，可将9项品质指标简化成3个主成分，其累积方差贡献率为74.5%，其中第一主成分因子的贡献率达到了 48.0%，第一主成分中因子载荷量大的品质指标是Zn、P、Ca、粗蛋白、γ-氨基丁酸、淀粉、Fe含量。综合评价筛选出10个营养品质较高的种质，分别为BJX152、BJX008、BJX149、BJX004、BJX003、BJX015、BJX005、藏青2000、BJX227、喜马拉22号。     

不同基因型大麦苗期对干旱胁迫的响应及HvPIPs基因表达特征分析

为研究不同基因型大麦苗期对干旱胁迫的响应,挖掘优质抗旱的大麦种质资源,以4份不同基因型大麦(2个高抗品种ZDM5430和ZDM5458,2个干旱敏感型品种7DCADA和IL-12)为材料,利用聚乙二醇(PEG-6000)模拟干旱条件,综合分析不同品种大麦的苗期抗旱性差异;并通过对不同品种根系进行PIPs实时定量分析,研究水通道蛋白AQPs基因与干旱胁迫之间的应答关系。结果表明,持续干旱胁迫会使大麦生长变缓,各处理苗长、根长、叶含水量和根含水量都呈下降趋势,其中,4个品种的大麦叶含水量在干旱胁迫14 d时下降最为显著,与对照相比,其降幅分别为29.14%、52.64%、21.67%和72.15%。干旱胁迫对大麦苗期干物质积累量和根冠比的影响较小,各处理间差异不显著;但持续干旱后,抗旱品种ZDM5430的根冠比呈上升趋势,在胁迫7、14、21 d时较CK分别增加了4.92%、42.86%和21.05%。随着胁迫时间的延长,大麦旗叶叶绿素含量和根系活力也呈下降趋势,其中ZDM5430和ZDM5458较对照组降幅较小,说明抗旱品种的形态指标、叶绿素含量及根系活力受干旱胁迫的影响较小。通过PIP...     

大小麦附加系的细胞学稳定性观察

对１Ａ，２Ａ，３Ａ，４Ａ，６Ａ及７Ａ等６个大小麦附加系的细胞学稳定性进行了观察。发现经两代繁殖后，其种子根尖细胞染色体有丢失现象。在减数分裂过程中，均观察到单价体、多价体、染色体断片、后期染色体落后、染色体桥及四分体微核等异常现象；经对四分体微核出现频率的统计，表明４Ａ，６Ａ及７Ａ等３个附加系在细胞学上相对较不稳定。     

Derivation and testing of a model to predict selection for fungicide resistance

A mathematical model was derived to predict selection for fungicide resistance in foliar pathogens of cereal crops. The model was tested against independent data from four field experiments quantifying selection for the G143A mutation conferring resistance to a quinone outside inhibitor (QoI) fungicide in powdery mildew (Blumeria graminis f.sp. hordei) on spring barley (Hordeum vulgare). Fungicide treatments with azoxystrobin differed in the total applied dose and spray number. For each treatment, we calculated the observed selection ratio as the ratio of the frequency of the resistant strain after the last and before the first spray. The model accurately predicted the variation in observed selection ratios with total applied fungicide dose and number of sprays for three of the four experiments. Underprediction of selection ratios in one experiment was attributed to the particularly late epidemic onset in that experiment. When the equation representing epidemic development was modified to account for the late epidemic, predicted and observed selection ratios at that site were in close agreement. On a scatter plot of observed selection ratios on predicted selection ratios, for all four experiments, the 1:1 line explained 89–92% of the variance in the mean of observed selection ratios. To our knowledge, this is the first fungicide resistance model for plant pathogens to be rigorously tested against field data. The model can be used with some degree of confidence, to identify anti‐resistance treatment strategies which are likely to be effective and would justify the resources required for experimental testing.