利用HPLC-ICP-MS方法测定中华绒螯蟹不同形态砷的残留量

采用高效液相色谱串联电感耦合等离子体质谱仪(HPLC-ICP-MS)测定中华绒螯蟹可食部位6种砷形态的残留量。经过前处理、提取和脱脂后,通过色谱柱、流动相、pH和流速等参数调节,筛选得到最优化的分离和检测条件。结果表明,6种砷形态的混合标准溶液在质量浓度0~50 μg/L范围时,均呈现良好的线性关系,其R²达到0.999,检测限为0.0060~0.0912 μg/kg,定量限为0.020~0.304 μg/kg。在样品中添加0.5、5、50 μg/L浓度水平,回收率良好,范围是70%~120%,相对标准偏差(RSD)均<5%,方法精密度与重复性均良好。本方法为中华绒螯蟹可食部位砷形态残留量建立了有效检测方法,为水产品砷残留快速检测提供技术支撑。     

水稻赖氨酸含量测定方法改进及其最优动态分类研究

赖氨酸是水稻营养成分中第一限制性氨基酸。为优化赖氨酸的测定方法,设置了本试验,通过超声振荡器取代普通振荡器,进行300W超声功率下酰化用时及染料结合反应用时的研究。结果表明,最佳酰化用时为15 min,染料结合用时为90 min。与国标法相比,优化后的方法酰化用时变长、染料结合用时变短,测定总用时明显缩短,但测定结果和试验精度更高。可见,采用超声波振荡代替传统振荡器振荡能改进赖氨酸的测定方法。另外,采用最优动态聚类法对赖氨酸含量进行分类,能防止人为分类的不确定性,做到分类方案的最优化。     

稻曲病菌交配型基因MAT1-2-1和MAT1-2-8的特性分析

 有性生殖在真菌的生活史和进化过程中具有重要作用,而交配型基因是控制有性生殖的关键因子。前期研究发现稻曲病菌(Villosiclava virens)MAT1-2型菌株中包含MAT1-2-1和MAT1-2-8两个交配型基因,但是它们如何调控稻曲病菌有性生殖依然不清楚。本文研究了它们在不同侵染和生长发育时期的表达模式和编码的蛋白结构特性。研究表明MAT1-2-1在侵染不同阶段一直下调表达;而MAT1-2-8在侵染早期(5 dpi)上调表达,在侵染后期下调表达。与营养菌丝阶段比较,MAT1-2-1和MAT1-2-8在有性发育过程菌核形成、菌核萌发、子座原基形成和子座成熟4个阶段的表达量都是下降的,在菌核形成阶段表达量最低。生物信息学分析显示MAT1-2-1和MAT1-2-8具有磷酸化位点,为非分泌蛋白,无明显的跨膜结构域。蛋白同源比对分析表明MAT1-2-1与香柱菌(Epichloë typhina)的MAT1-2-1同源性最高,而MAT1-2-8与绿僵菌(Metarhizium)的MBR_08192蛋白同源性最高。进一步研究发现MAT1-2-1和MAT1-2-8能够互作,并分别主要定位在细胞核和细胞基质中。通过质谱技术鉴定到MAT1-2-1的一些候选互作蛋白,如假定Ran交换因子Prp20/Pim1(KDB12229.1)、假定rRNA处理蛋白Ebp2(KDB12923.1)及组蛋白H1(KDB12711.1)等。因此,以上结果为研究稻曲病菌交配型基因MAT1-2-1和MAT1-2-8调控有性生殖的生物学功能奠定了基础。     

OsHemA gene,encoding glutamyl-tRNA reductase(GluTR) is essential for chlorophyll biosynthesis in rice(Oryza sativa)

Chlorophyll(Chl) biosynthesis is essential for photosynthesis and plant growth. Glutamyl-tRNA reductase(GluTR) catalyzes glutamyl-tRNA into glutamate-1-semialdehyde(GSA) and initiates the chlorophyll biosynthesis. Even though the main role of GluTR has been established, the effects caused by natural variations in its corresponding gene remain largely unknown. Here, we characterized a spontaneous mutant in paddy field with Chl biosynthesis deficiency, designated as cbd1. With intact thylakoid lamellar structure, the cbd1 plant showed light green leaves and reduced Chl and carotenoids(Cars) content significantly compared to the wild type. By map-based gene cloning, the mutation was restricted within a 57-kb region on chromosome 10, in which an mPingA miniature inverted-repeat transposable element(MITE) inserted in the promoter region of OsHemA gene. Both leaf color and the pigment contents in cbd1 were recovered in a complementation test, confirming OsHemA was responsible for the mutant phenotype. OsHemA was uniquely predicted to encode GluTR and its expression level was dramatically repressed in cbd1. Transient transformation in protoplasts demonstrated that GluTR localized in chloroplasts and a signal peptide exists in its N-terminus. A majority of Chl biosynthesis genes, except for POR and CHLG, were down-regulated synchronously by the repression of OsHemA, suggesting that an attenuation occurred in the Chl biosynthesis pathway. Interestingly, we found major agronomic traits involved in rice yield were statistically unaffected, except for the number of full grains per panicle was increased in cbd1. Collectively, OsHemA plays an essential role in Chl biosynthesis in rice and its weak allele can adjust leaf color and Chls content without compromise to rice yield.     

不同氮效率油菜种质苗期氮吸收转运与利用差异研究

氮效率研究是油菜营养性状遗传改良的前提,为探究不同氮效率油菜种质苗期氮吸收、转运和利用的异同,以2个氮效率差异油菜种质H6（氮高效）和L18（氮低效）为供试材料,利用水培营养液设置正常氮（CK）和低氮（LN）2个氮浓度处理,培养14d后检测植株氮含量,计算氮累积量、氮转运系数和氮利用效率。结果显示,不同氮浓度处理对油菜苗期氮的吸收、转运和利用效率影响的差异达到极显著水平（P＜0.01）。与CK相比,LN处理的油菜生物量、氮含量和氮累积量均显著降低,而氮利用效率和根冠比显著提高;氮高效油菜种质H6的生物量、氮累积量、氮转运系数和氮生理利用效率均显著大于氮低效种质L18,分别为L18的2.07、1.42、3.23和1.56倍。从氮的吸收、转运和利用3个方面探讨了低氮胁迫处理下油菜种质苗期氮效率差异的原因,对深入开展油菜氮高效机理研究及油菜品种氮效率改良具有一定的指导意义。     

The integration and insertion site of GbVe1 gene in the genome of transgenic cotton (Gossypium hirsutum)

[Objective] The aim of this study was to obtain the flanking sequences of T-DNA in the transgenic cotton containing a GbVe1 over-expression cassette. [Method] The T-DNA insertion copy number in the transgenic GbVe1 cotton was analyzed by southern blot. Flanking sequences of the transgenic lines with putative single T-DNA insertion copy were obtained using high-efficiency Thermal asymmetric interlaced polymerase chain reaction (hiTAIL-PCR). The T-DNA insertion sites were further confirmed by PCR with specific primers. [Result] RB-flanking sequences (119-1 018 bp) and LB-flanking sequences (243-516 bp) were obtained from three transgenic lines with low copy number of T-DNA insertion. The AT content was more than 63% in these flanking sequences. A same single insertion site in the intron of Gohir.D01G157600.1 was found in the two transgenic lines 7/100826-152 and 12/100826-393, while two separated insertion sites, one also in the intron of Gohir.-D01G157600.1 and the other in the intergenic region of A12 chromosome, were found in the transgenic line 1/w-ch14. A deletion of 21 bp was found in the insertion site in the intron of Gohir.D01G157600.1. The T-DNA insertion in the intron of Gohir.D01G157600.1 was further confirmed by the specific PCR. [Conclusion] The flanking sequences of T-DNA in the transgenic GbVe1 cotton were obtained and the specific transformation event in the intron of Gohir.D01G157600.1 was further confirmed by PCR.     

我国蔬菜作物耐低温性研究进展

低温胁迫是蔬菜生产过程中的主要逆境因子，对蔬菜作物的生长发育、商品产量形成影响极大。本文综述了蔬菜作物耐低温性的遗传模型、生理调节和分子机制，展望了蔬菜作物耐低温性的研究前景。     

稻曲病菌致病机制研究进展

 稻曲病是一种水稻穗部病害,在世界各水稻主产区均有发生,已成为世界性病害,在我国也属于水稻主要病害之一。稻曲病的发生严重影响水稻产量和稻米品质。研究其病原菌致病机制,对找寻防治药物的特异性靶标具有重要作用,可为抗病分子育种提供新的思路,为稻曲病菌防治药剂研发提供新的靶标。本文综述了稻曲病菌侵染过程、致病相关基因功能研究、水稻响应稻曲病菌侵染等方面的研究进展,提出进一步研究的策略。现有研究表明,稻曲病菌能成功侵染水稻孕穗期雄蕊的花丝引起发病。病原菌可通过抑制多个水稻免疫途径实现成功侵染;还可模仿胚珠受精激活水稻灌浆、糖代谢等途径为稻曲球的形成提供营养物质;全基因组测序完成和基因编辑技术在基因敲除中的成功应用,为稻曲病菌功能基因研究提供了很好的基础。稻曲病菌独特的侵染过程和营养利用机制是未来研究的重要方向。     

不同类型水稻品种产量和氮素吸收利用对大气CO2浓度升高响应的差异

目的 探明不同类型水稻品种产量和氮素吸收利用对FACE(大气CO₂浓度增高)响应的差异。方法 以常规粳稻、杂交籼稻、常规籼稻共6个品种为供试材料,研究FACE对不同类型水稻产量、氮素吸收利用的影响。结果 1）FACE处理极显著提高了水稻产量,平均增加24.17%, 常规籼稻增幅最大,FACE和对照均以杂交籼稻最高;2）FACE处理显著增加了单位面积穗数,常规粳稻增幅最大,并显著增加了杂交籼稻和常规籼稻每穗粒数;3）FACE处理显著提高了成熟期吸氮量和氮素籽粒生产效率,成熟期吸氮量平均增加21.23%,杂交籼稻增幅最大, FACE和对照均以常规籼稻最高;氮素籽粒生产效率平均增加7.33%,杂交籼稻增幅最大,FACE和对照均以杂交籼稻最高。成熟期吸氮量对产量促进作用略大于成熟期氮素籽粒生产效率;4）FACE处理降低了植株含氮率,成熟期平均下降0.105个百分点,常规粳稻降幅最大。FACE处理极显著提高植株干物质量,成熟期平均增加23.95%,常规籼稻增幅最大;FACE处理显著提高常规籼稻和杂交籼稻成熟期单穗吸氮量,分别增加10.79%、13.93%,但常规粳稻下降了9.60%;FACE处理显著提高了成熟期群体吸氮强度,平均增加22.29%,杂交籼稻增幅最大。FACE处理对水稻全生育期天数无显著影响;FACE处理显著提高茎鞘、叶片、穗各器官吸氮量,叶片增幅最大,平均增加51.86%,杂交籼稻增幅最大;FACE处理显著提高了不同生育阶段吸氮量,抽穗-成熟阶段增幅最大,平均增加108.90%,杂交籼稻增幅最大;5）植株干物质量、单穗吸氮量、吸氮强度、穗吸氮量、抽穗-成熟阶段吸氮量对成熟期总吸氮量的促进作用分别大于植株含氮率、单位面积穗数、生育天数、茎鞘叶吸氮量、移栽-分蘖和分蘖-抽穗阶段吸氮量;6）FACE处理显著提高了氮肥偏生产力,降低了每百千克籽粒需氮量,前者平均增加24.16%,常规籼稻增加最多;后者平均降低4.7%,常规籼稻降幅最大。结论 FACE处理可显著提高水稻产量和氮素吸收利用效率,但品种间差异较大。     

Wx^mp基因背景下可溶性淀粉合成酶基因SSⅡa和去分支酶基因PUL对水稻蒸煮食味品质的影响

【目的】分析在同一主效基因(Wx^mp)背景下可溶性淀粉合成酶基因SSⅡa和去分支酶基因PUL对稻米蒸煮食味品质的影响,以期为水稻品质遗传改良提供依据。【方法】选择在SSⅡa和PUL存在多态性而其他淀粉合成酶相关基因没有多态性的半糯品系宁0145和粳稻品种武运粳21进行杂交,获得F2群体与F3株系。利用分子标记,选择含有Wx^mp基因的F2单株与F3株系,将这些F2单株与F3株系分成SSⅡa^nPUL^n、SSⅡa^nPUL^w、SSⅡa^wPUL^n和SSⅡa^wPUL^w4种基因型(n和w分别表示该基因来源于宁0145和武运粳21),分析不同基因型蒸煮食味品质性状的差异,探讨同一Wxmp基因背景下不同SSⅡa和PUL等位基因对蒸煮食味品质性状的影响。【结果】不同基因型间蒸煮食味品质性状均存在显著差异,来源于武运粳21的SSⅡa^w基因和PUL^w基因分别使直链淀粉含量增加0.29%~1.00%和0.62%~1.18%,且PUL的效应大于SSⅡa,两者间存在互作效应。SSⅡa^w基因和PUL^w基因降低胶稠度和崩解值,提高了热浆黏度、冷胶黏度、消减值和回复值,对糊化温度、峰值黏度和峰值时间的作用较小。【结论】明确了Wx^mp背景下SSⅡa和PUL基因对稻米蒸煮食味品质的遗传效应,该研究结果为SSⅡa和PUL基因的分子标记辅助选择改良稻米品质提供了理论依据。