hrpZ_(Psta)在转基因大豆中定量表达与疫霉根腐病和灰斑病抗性相关研究

以T5转hrpZPsta基因大豆JN29-705-15和JL30-187为材料,利用实时荧光定量PCR技术(Quantitative Real Time PCR,qRT-PCR)检测了目标基因在转基因大豆不同组织中的表达量,分别利用下胚轴侵染法和叶面喷施法鉴定疫霉根腐病抗性和灰斑病抗性,并分析目的基因表达量与疫霉根腐病和灰斑病抗性的相关性。结果表明:hrpZPsta基因在大豆的叶、茎、根、籽粒中均有表达,二个株系平均相对表达量分别为8.2/6.1、0.9/0.7、6.5/4.6和0.8/0.7;T5转hrpZPsta基因大豆抗疫霉根腐病和灰斑病能力与野生型相比均有所提高,JN29-705-15对疫霉根腐病抗性从感病提高到中抗,而JL30-187从中抗提高到抗病;hrpZPsta基因在叶中的表达量也与抗灰斑病能力呈正相关,与病情级别呈极显著负相关。试验结果初步证明了外源基因hrp ZPsta在大豆植株中的表达量与受体植株对疫霉根腐病和灰斑病抗性存在一定的相关性。     

Effects of Glyphosate on Soil Salinization and Alkalization in Cotton Fields

We analyzed the effects of glyphosate to the index of salinization and alkalization by simulation in soils collected from cotton fields in Anyang, Henan and Anqing, Anhui, China, in 2014. The results showed that application of glyphosate changed electrical conductivity (EC), total alkalinity (TAL), exchangeable sodium percentage (ESP), sodium adsorption ratio (SAR) and pH of soil in both regions，compared with the blank control. With increased frequency of glyphosate application and increased concentration applied, all indices increased in the Anqing cotton fields soil which were treated with concentrations of 2.5-20 g·L^－1 . However, EC, TAL and ESP increased dynamically in Anyang soil to which high concentrations of 20 g·L^－1 were applied only four consecutive times. After different glyphosate treatments, the soil pH value showed an upward trend but smaller changes. Therefore, after repeated use of glyphosate to soils of two different region soils, although the changes of all indices were within the range of non-saline and non-sodic soils, the soils affected by salt in the glyphosate aqueous solution tended towards saline-sodic soils.     

棉花SSR-PCR过程中聚丙烯酰胺凝胶电泳操作常见问题的解决与优化

在大批量聚丙烯酰胺凝胶电泳过程中,难免会出现条带空白、弯曲或者拖尾等各种异常现象。本文以棉花DNA为例,综述了多年实验操作中遇到的突发情况及解决方法,为广大科研工作者提供参考。     

褚橙龙陵基地柑橘叶片DRIS图解法和指数法综合营养诊断分析

[目的]开展基于综合营养诊断施肥法(Diagnosis and Recommendation Integrate System,DRIS)的冰糖橙种植基地柑橘营养诊断,为柑橘园的营养诊断和平衡施肥提供参考依据.[方法]以云南褚橙龙陵基地500 ha冰糖橙为研究对象(柑橘园随机划分为8个区域),通过测定成熟秋梢叶片中氮(N)、磷(P)、钾(K)、钙(Ca)、镁(Mg)、铜(Cu)、锌(Zn)和铁(Fe)矿质营养的含量,运用DRIS图解法求得冰糖橙叶片各营养元素诊断参数最适比范围,依据DRIS指数法求得各区域冰糖橙需肥紧迫程度排序及养分不平衡指数(NII),初步制定冰糖橙树体营养元素DRIS指数分级标准.[结果]整个园区高产园(>22500 kg/ha)的各营养元素含量大多高于低产园(≤22500 kg/ha),高产园和低产园间N、Fe、N/Fe、P/Fe、Ca/Mg、Ca/Fe、Mg/Zn差异显著(P<0.05)或极显著(P<0.01).当N/P在2.27~2.57,N/K在1.58~1.91,P/K在0.66~0.79时冰糖橙叶片N、P和K养分含量平衡;当Ca/Mg、Ca/Fe、Mg/Fe、Cu/Zn、Cu/Fe和Zn/Fe分别在17.01~19.09、0.21~0.26、0.012~0.015、2.77~4.60、0.30~0.43和0.09~0.12时,冰糖橙叶片Ca、Mg、Fe、Zn和Cu养分处于平衡状态.综合诊断显示冰糖橙叶片养分需求顺序为Mg>N>K>P>Ca>Cu>Zn>Fe,不同区域冰糖橙养分需求顺序不同,高产园冰糖橙相对缺乏的养分有N、P,低产园相对缺乏的养分有N、Mg,相比之下,低产园营养比例失衡情况较严重.依据高产园叶片矿质营养元素含量范围确定冰糖橙叶片矿质营养元素的适宜值为N(11.66~13.30 g/kg)、P(4.74~5.61 g/kg)、K(6.23~8.23 g/kg)、Ca(13.67~19.33 g/kg)、Mg(0.86~0.97 g/kg)、Cu(14.49~37.71 mg/kg)、Zn(6.38~8.02 mg/kg)和Fe(55.91~85.74 mg/kg).[结论]根据DRIS诊断,褚橙龙陵基地冰糖橙园普遍存在N、P、K、Ca、Mg缺乏,Cu、Zn、Fe相对过剩问题.DRIS图解法和指标法综合诊断既可求得各元素最适比范围和需肥紧迫程度,同时,参照冰糖橙叶片养分含量分级标准能判断树体营养元素的平衡状况,可用于生产上指导下一年合理施肥.     

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.     

Pathogen Analysis of Goat's Respiratory Diseases

To investigate the pathogens of goat's respiratory disease from a goat farm in Guangxi province, the epidemiological investigation, clinic observation, pathological examination, bacterium isolation and identification, biochemical test, PCR and animal experiment test were conducted.The preventive and control measures were taken on the basis of pathogen epidemiological characteristics and drug sensitive test results.The results showed that Mycoplasma, influenza virus and parasite were negative by isolation and culture or PCR, however, two strains of gram-negative bacteria named MS1 and MS2 were isolated from lung.MS1 was identified as Serratia marcescens by biochemical test and 16S rRNA sequencing, which shared 99% nucleotide homology with other Serratia marcescens in GenBank.And MS2 was identified as E.coil by the same methods that shared 99% nucleotide homology with other E.coli in GenBank.Animal experiment showed that two strains could cause death in mice.The drug sensitive tests showed two strains were highly sensitive to spectinomycin, amikacin, kanamicin and neomycin.Kanamycin and dexamethasone were used to treat the sick goats, and achieved good results.     

不同叶龄蘖、穗氮肥组合对粳稻产量及氮素利用的影响

以主茎叶片数不同的粳稻品种吉粳88 (14片)、沈农265 (15片)和沈农1401 (16片)为试材,采用大田筒栽方式,在总施氮量225kghm–2及轻简施肥(基肥、蘖肥、穗肥)模式基础上,设置基蘖肥∶穗肥6∶4和8∶2两种施肥比例,并分设不同源、库叶龄期施氮组合即不同叶龄蘖、穗肥精确施氮组合。分析了不同源库期氮肥运筹模式对水稻农艺性状、产量及氮素利用特性的影响。结果表明:(1)在有效穗数、分化颖花数、产量和氮素利用率方面,吉粳88、沈农265、沈农1401不同氮肥运筹下最佳蘖、穗肥叶龄组合均为6∶4显著高于8∶2。(2)不同氮肥运筹下,吉粳88在8叶(叶龄指数57.1%)、沈农265在9叶(叶龄指数60.0%)、沈农1401在10叶(叶龄指数62.5%)时,即叶龄指数在60%左右时,施用蘖肥效果最佳,最终穗数最多,对保蘖起主要作用;吉粳88在11叶(叶龄指数78.6%)、沈农265在12叶(叶龄指数80.0%)、沈农1401在13叶(叶龄指数81.3%)时,即叶龄指数在80%左右时,施用穗肥效果最佳,最终穗粒数最多,对促花起主要作用。(3)吉粳88-6∶4 (8, 11),沈农265-6∶4 (9, 12),沈农1401-6∶4 (10, 13) 3组处理,在产量、氮素积累量、氮素吸收利用率、农学利用率及偏生产力等方面,显著高于同品种不同叶龄蘖、穗氮肥组合中的其他处理。因此,适当延迟蘖肥施用叶龄期(叶龄指数60%左右)、提前穗肥施用叶龄期(叶龄指数80%左右)同时增加穗肥施用比例,既可以显著提高氮素积累量、氮素吸收利用率、农学利用率及偏生产力,又能显著促进成穗率的提高和颖花数的分化,达到保蘖促花的双重作用,实现优源、扩库、充实的目标,从而获得高产。     

质量控制图在水产饲料氨基酸分析中的应用

通过对实验室自制标准物料-豆粕样品中氨基酸含量的多次测定,建立X-S质量控制图,开展水产饲料氨基酸分析日常质量控制。试验按照GB/T 18246-2000,通过微波水解-离子交换-柱后衍生-紫外检测法对豆粕样品氨基酸含量进行连续测定,并以FAPAS奶粉标准物质验证,利用偏度-峰度检验法对离群值进行判断,建立赖氨酸、甘氨酸、氨基酸总量质量控制图。结果显示,质量控制图中豆粕质控样赖氨酸、甘氨酸、氨基酸总量测定的RSD值均小于4%,FAPAS奶粉标准物质测定值落在参考范围内,建立的质量控制图能对日常氨基酸检测结果进行有效监控。结果表明,所建立氨基酸质量控制图直观有效,能对过程存在的异常因素起到预警作用,并可节约成本。     

Effects of Adding Mannan Oligosaccharides to Diets with Different Concentrate to Forage Ratios on Ruminal Fermentation of Sheep in vitro

The test was aimed at determining the effects of adding MOS to diets with different concentrate to forage ratios on ruminal fermentation of sheep in vitro.The 4×6 two-factor experimental design was chosen.There were 4 diets with different concentrate to forage ratios(20:80, 30:70, 40:60 and 50:50) and added to 6 dosages MOS(0, 0.4%, 0.8%, 1.2%, 1.6% and 2.0%), respectively.The method of gas production in vitro was used to measure the total gas and CH₄ production, IVDMD, IVCPD and IVOMD after cultured for 3, 6, 9, 12 and 24 h.The results showed that the gas(except 12 and 24 h) and CH₄ production(except 9 h) were not impacted by the two factors(P>0.05).The IVDMD, IVCPD and IVOMD were significantly affected by concentrate to forage ratios(P<0.05), while the IVCPD at 24 h was significantly affected by MOS(P<0.05).Meanwhile, all the indexes were not influenced by the interaction of two factors(P>0.05).The IVDMD and IVOMD increased with the rising of concentrate approximately, while the IVCPD decreased with the rising of MOS roughly.The biggest associative effect was gained in 30:70 concentrate to forage diet when adding 1.2% MOS.     

小麦族的基因组显性及其育种学意义

小麦族包含大量由不同基因组组成的异源多倍体物种。同一个异源多倍体物种的不同基因组可能对表型性状产生非对称性贡献,例如小麦属多倍体物种的形态分类特性,更像其A基因组供体物种,这种现象称为A基因组显性。由于基因组显性,小麦族形成了以A、D、U、St为轴心(显性)基因组的异源多倍体物种簇。异源多倍体物种的基因组显性可能与其进化适应优势的形成有关。在育种方面,基因组显性影响多倍体新作物开发及小麦-外源染色体易位设计。