全长cDNA文库的构建方法

全长cDNA文库的构建是进行功能基因组研究的一种经济、快速、有效的途径,克服了传统cDNA文库的缺点,节省了基因克隆和功能鉴定的时间,促进了功能基因组的研究进程。目前有6种构建全长cDNA文库的方法,包括Oligo-Capping、CAPture、SMART、CAP-trapper、CapSelect、CAP-jumping。这几种方法在起始材料用量、文库构建技术、实验操作复杂程度和文库构建质量等方面构存在不同程度的优缺点,但综合比较而言,SMART和CAP-trapper这两种方法比较有实际应用价值。SMART法适于简单、快速地构建一般质量的cDNA文库,如果采用Little-cycle SMART和Large-size SMART这两种改进技术,所建文库质量也非常好;CAP-trapper法虽然对实验技术要求较高,实验过程复杂,但所建文库全长率高,冗余性低,建库效率高,是目前构建高质量文库最好的方法。     

优良水稻染色体片段代换系Z746的鉴定及重要农艺性状QTL定位及其验证

粒型、株高及穗部组成性状与产量形成密切相关,是水稻重要农艺性状,但遗传基础复杂。染色体片段代换系是用于复杂性状遗传研究的良好材料。本研究鉴定了一个以日本晴为受体、西恢18为供体亲本的水稻优良染色体片段代换系Z746。Z746携带来自西恢18的7个代换片段,平均代换长度为3.99 Mb,其株高、粒长和穗部性状均与受体存在显著差异。进一步通过日本晴与Z746杂交构建的次级F2群体共检测到36个相关QTL,分布于2号、3号、4号、6号和11号染色体。其中5个可能与已克隆基因等位,如qPH3-1等, 8个可被多次检出,表明这些是遗传稳定的主效QTL。Z746的粒长主要由4个QTL(qGL3、qGL4、qGL2、qGL6)控制,其中qGL3和qGL4对粒长变异的贡献率分别为60.28%和27.47%。株高由5个QTL控制,穗长由4个QTL控制,每穗粒数由2个QTL控制,千粒重由2个QTL控制。然后以MAS在F3共选出8个单片段代换系,并以此在F4进行了相关QTL验证,共有24个QTL可被8个单片段代换系(SSSL)检出,重复检出率为66.7%,表明这些QTL遗传稳定。本研究为目的 QTL的进一步...     

氮响应转录因子Zm NLP5影响玉米根系生长的功能研究

研究作物氮高效机制并挖掘利用其中重要的调控基因,对我国农业绿色发展具有重要意义。前期我们发现氮响应转录因子ZmNLP5调控亚硝酸还原酶基因的表达,对玉米氮素吸收利用具有促进作用,但其调控机制尚未明确。基于此,本研究以ZmNLP5基因突变体(zmnlp5)和野生型(WT)植株为研究材料,深入解析ZmNLP5影响玉米氮素吸收利用的生理机制。足氮(sufficient nitrogen, SN)和低氮(deficient nitrogen, DN)条件下研究材料表型分析发现, DN条件下相对于WT植株, zmnlp5植株根长显著降低。根部不同区域ZmNLP5表达量分析发现, ZmNLP5主要在根尖区域表达。不同亚硝酸盐浓度处理下,研究材料的根长和根尖区域亚硝酸盐含量分析发现,当亚硝酸盐的浓度高于2mmol L~(-1)时,zmnlp5突变体根长相对于WT显著降低,zmnlp5植株根尖区域亚硝酸盐积累量显著高于WT。SN和DN条件下研究材料根部亚硝酸盐含量检测发现, DN条件zmnlp5植株根尖区域所积累的亚硝酸盐也显著高于WT。综上所述,转录因子ZmNLP5在玉米植株响应低氮环境根部伸长生长中发挥重要功能,该结果为玉米氮高效育种提供了一定的理论基础。     

基于叶片SPAD的棉花氮营养指数估算研究

为了探究不同施氮处理对棉花叶片SPAD值、生物量和氮营养指数(NNI)的影响,通过两年田间试验,设置5个施氮水平(即0、120、240、360、480 kg/hm2),开展棉花叶片SPAD值、氮素吸收分配规律和生物量变化的研究.研究结果发现:棉叶SPAD值随施氮量增加呈增加趋势,在N4处理下获得最大值;建立棉花氮浓度稀...     

Quantitative trait loci (QTL) mapping of silique length and petal colour in Brassica rapa

Recombinant inbred lines (RILs) derived from a cross between Brassica rapa L. cv. ‘Sampad’, and an inbred line 3‐0026.027 was used to map the loci controlling silique length and petal colour. The RILs were evaluated under four environments. Variation for silique length in the RILs ranged from normal, such as ‘Sampad’, to short silique, such as 3‐0026.027. Three QTL, SLA3, SLA5 and SLA7, were detected on the linkage groups A3, A5 and A7, respectively. These QTL explained 36.0 to 42.3% total phenotypic variance in the individual environments and collectively 32.5% phenotypic variance. No additive × additive epistatic interaction was detected between the three QTL. Moreover, no QTL × environment interaction was detected in any of the four environments. The number of loci for silique length detected based on QTL mapping agrees well with the results from segregation analysis of the RILs. In case of petal colour, a single locus governing this trait was detected on the linkage group A2.     

甘蓝型油菜角果长度的主+多基因混合遗传模型

角果是油菜产量构成要素中重要的组成部分。本文以长角果品种中双11和短角果材料10D130为亲本配制杂交组合,采用主基因+多基因混合遗传模型分析方法对该组合6世代遗传群体(P₁、P₂、F₁、BCP₁、BCP₂和F₂)的果身长、角果长和果喙长进行遗传分析。结果表明,该组合的3个角果性状均呈连续分布,其中,果身长最适遗传模型为E-0 (2对加性-显性-上位性主基因+加性-显性-上位性多基因模型),2对主基因加性效应值分别是1.75和–0.06,显性效应值分别是–0.59和–0.86,主基因遗传率在BCP₁、BCP₂和F₂中分别是51.10%、74.23%和66.93%,多基因遗传率分别为29.16%、17.11%和23.96%。角果长的最适遗传模型为E-1 (2对加性-显性-上位性主基因+加性-显性多基因模型),其中,第1对主基因加性效应为0.34,显性效应为–0.81,第2对主基因加性效应为0.34,显性效应为–0.47,主基因遗传率在BCP₁、BCP₂和F₂中分别是47.63%、68.51%和79.45%,多基因遗传率分别为29.40%、20.89%和12.47%。果喙长的最适遗传模型为E-3模型(2对加性主基因+加-显多基因遗传模型),2对主基因加性效应值分别是0.2和–0.2,主基因遗传率在BCP₁、BCP₂和F₂中分别是33.71%、72.75%和52.25%,多基因遗传率分别为40.08%、5.37%和27.60%。     

种带真菌对结缕草种子发芽和活力的影响

采用孢子悬浮液法将从结缕草上分离得到的种带真菌菌种进行回接，测试不同种带真菌对结缕草发芽和活力的影响。研究结果表明：多数种带真菌显著降低了种子的发芽率和活力指数，抑制了种苗的生长，使死种率显著上升，其中，不等弯孢（Curvularia inaequalis）、禾生皮司霉（Pithmyces graminicola）、橡胶平脐蠕（Bipolaris heveae）、嘴突突脐蠕孢（Exerohilum rostratum）、白粉寄生菌（Ampelomyces sp．）、狗尾草平脐蠕孢（B．setariae）、茎点霉（Phoma sp．）、木贼镰刀菌（Fusarium equiseti）等对结缕草种子和种苗的危害最为严重。     

Spatial and temporal distribution of the root system and root nutrient content of an established Miscanthus crop

Although a high biomass yield is obtained from established Miscanthus crops, previous studies have shown that fertilizer requirements are relatively low. As little information on the role of the Miscanthus roots in nutrient acquisition is available, a study was conducted to gather data on the Miscanthus root system and root nutrient content. Therefore in 1992, the root distribution pattern of an established Miscanthus crop was measured in field trials using the trench profile and the auger methods. Also, in 1994/1995, seasonal changes in root length density (RLD) and root nutrient content were monitored three times during the vegetation period.

The trench profile method showed that roots were present to the maximum depth measured of 250 cm. The top soil (0–30 cm) contained 28% of root biomass, while nearly half of the total roots were present in soil layers deeper than 90 cm. Using the auger method, we found that RLD values in the topsoil decreased with increasing distance from the centre of the plants. Below 30 cm, RLD decreased markedly, and differences in root length in the soil between plants were less pronounced. The total root dry weight down to 180 cm tended to increase from May 1994 (10.6 t ha⁻¹) to November 1994 (13.9 t ha⁻¹) and then decreased again until March 1995 (11.5 t ha⁻¹). Nutrient concentrations in the roots decreased with increasing depth. The concentrations of N (0.7–1.4%) and K (0.6–1.2%) were clearly higher than those of P (0.06–0.17%). The mean values for N, P and K contents of the roots of all three sampling dates in 1994/1995 were 109.2 kg N ha⁻¹, 10.6 kg P ha⁻¹ and 92.5 kg K ha⁻¹.

Although our results showed that RLD values for Miscanthus in the topsoil are lower than for annual crops, the greater rooting depth and the higher RLD of Miscanthus in the subsoil mean that nutrient uptake from the subsoil is potentially greater. This enables Miscanthus crops to overcome periods of low nutrient (and water) availability especially during periods of rapid above-ground biomass growth.     

Separating N2O production and consumption in intact agricultural soil cores at different moisture contents and depths

Agricultural soils are a major source of the potent greenhouse gas and ozone depleting substance, N₂O. To implement management practices that minimize microbial N₂O production and maximize its consumption (i.e., complete denitrification), we must understand the interplay between simultaneously occurring biological and physical processes, especially how this changes with soil depth. Meaningfully disentangling of these processes is challenging and typical N₂O flux measurement techniques provide little insight into subsurface mechanisms. In addition, denitrification studies are often conducted on sieved soil in altered O₂ environments which relate poorly to in situ field conditions. Here, we developed a novel incubation system with headspaces both above and below the soil cores and field-relevant O₂ concentrations to better represent in situ conditions. We incubated intact sandy clay loam textured agricultural topsoil (0–10 cm) and subsoil (50–60 cm) cores for 3–4 days at 50% and 70% water-filled pore space, respectively. ¹⁵N-N₂O pool dilution and an SF₆ tracer were injected below the cores to determine the relative diffusivity and the net N₂O emission and gross N₂O emission and consumption fluxes. The relationship between calculated fluxes from the below and above soil core headspaces confirmed that the system performed well. Relative diffusivity did not vary with depth, likely due to the preservation of preferential flow pathways in the intact cores. Gross N₂O emission and uptake also did not differ with depth but were higher in the drier cores, contrary to expectation. We speculate this was due to aerobic denitrification being the primary N₂O consuming process and simultaneously occurring denitrification and nitrification both producing N₂O in the drier cores. We provide further evidence of substantial N₂O consumption in drier soil but without net negative N₂O emissions. The results from this study are important for the future application of the ¹⁵N-N₂O pool dilution method and N budgeting and modelling, as required for improving management to minimize N₂O losses.