海马齿SpSCL1基因启动子克隆及序列分析

GRAS蛋白是植物特有的一类蛋白家族,在植物的生长发育方面具有重要作用.根据前期分离的海马齿SpSCL1基因的5'端序列,设计特异引物进行Genome Walking,从海马齿基因组DNA中克隆了SpSCL1基因上游的1 254 bp片段.序列分析表明,该序列包含681 bp的启动子调控序列及573 bp的基因编码序列,启动子调控序中除含有典型的真核生物核心启动子区域(-60～-10bp)外,还含有多个TATA-box、CAAT-box等启动子元件以及多种与脱水、耐盐和光响应相关的顺式作用元件.结果预示海马齿SpSCL1可能在抗旱、耐盐以及光敏色素信号转导途径中发挥重要作用.     

甘蔗乙烯受体Sc-ERS启动子的克隆与序列分析

【目的】克隆甘蔗乙烯受体Sc-ERS基因启动子序列,分析Sc-ERS基因启动子序列的作用元件。【方法】采用基于热不对称交错式PCR原理的染色体步移技术,从甘蔗ROC22基因组DNA中克隆Sc-ERS基因启动子序列,利用启动子预测软件PlantCARE和PLACE在线工具对Sc-ERS启动子序列的作用元件进行预测。【结果】克隆获得Sc-ERS启动子序列1410 bp, 该序列与玉米ERS25、ERS14核苷酸同源性分别为82%和80%。PlantCARE 在线分析结果表明,该序列具有启动子基本作用元件TATA-BOX和CAAT-BOX,还含有参与光响应元件、干旱诱导MYB 结合位点、茉莉酸甲酯响应元件、水杨酸响应元件、胚乳表达顺式调控元件、热胁迫响应元件等。【结论】从甘蔗基因组DNA中克隆获得乙烯受体Sc-ERS基因上游1410 bp的启动子序列,该序列含有多个特异性调控元件,推测Sc-ERS基因的表达可能受生理周期、激素、干旱、光照等因素调控。     

甘蔗乙烯受体Sc-ERS启动子的克隆与序列分析

【目的】克隆甘蔗乙烯受体Sc-ERS基因启动子序列,分析Sc-ERS基因启动子序列的作用元件。【方法】采用基于热不对称交错式PCR原理的染色体步移技术,从甘蔗ROC22基因组DNA中克隆Sc-ERS基因启动子序列,利用启动子预测软件PlantCARE和PLACE在线工具对Se-ERS启动子序列的作用元件进行预测。【结果】克隆获得sc．ERS启动子序列1410bp,该序列与玉米ERS25、ERS14核苷酸同源性分别为82％和80％。PlantCARE在线分析结果表明,该序列具有启动子基本作用元件TATA．BOX和CAAT—BOX,还含有参与光响应元件、干旱诱导MYB结合位点、茉莉酸甲酯响应元件、水杨酸响应元件、胚乳表达顺式调控元件、热胁迫响应元件等。【结论】从甘蔗基因组DNA中克隆获得乙烯受体Sc-ERS基因上游1410bp的启动子序列,该序列含有多个特异性调控元件,推测sc．ERS;~因的表达可能受生理周期、激素、干旱、光照等因素调控。     

谷子GATA基因家族的鉴定及表达分析

本研究从谷子基因组水平鉴定出33个SiGATA成员,命名为SiGATA1～SiGA TA33.利用生物信息学方法对谷子SiGATA家族的系统发育、基因结构、染色体分布、保守基序、蛋白三级结构和顺式调控元件进行预测和分析.结果 表明,33个SiGATA转录因子被划分为3组,不均匀的分布在8条染色体上,多数含有CX2CX1...     

Research on Yucca schidigera extract feeding on the rumen ecology, protozoal populations and serum chemistries of sheep

In a completely randomized block design experiment, 16 ruminally cannulated male sheep with body weights of (40 ± 2.1) kg were fed twice daily (8:00 and 16:00) with concentrate and forage (50:50 on dry matter (DM) basis). Dietary treatments were supplemented with intraruminal doses of powdered Yucca schidigera extract (YSE) at the levels of 0 (control), 100, 200 and 300 mg · kg⁻¹. On days of 15, 16 and 17 after feeding, ruminal content was sampled at 0, 2, 4, 6 and 8 h after dosing (8:00), and blood samples were collected at the end of experiment (the days 18 and 19 after feeding). Results showed that the treatment groups’ acidity was not affected (P = 0.13) by YSE. Comparing to the control, the ruminal propionate concentration was increased by YSE addition in a dose-dependent manner by up to 29.8% (P < 0.05), and the acetic concentration was decreased by up to 17.5% (P < 0.05). The ruminal ammonia concentration 2 hours after feeding was higher (P < 0.05) in sheep fed without YSE (increased by 17.57 mg· 100 mL⁻¹) than those fed with YSE at 200 mg · kg⁻¹ (6.77 mg · 100 mL⁻¹ increase in NH₃) and at 300 mg · kg⁻¹ (6.50 mg· 100 mL⁻¹ increase in NH₃). Protozoal populations in the rumen were lower (P < 0.05) with the YSE feeding dose at 300 mg · kg⁻¹ than the control. The serum chemistries were not different among treatments (P > 0.05) and were within the normal physiological ranges for sheep 19 days after feeding. The study indicated that 200 mg· kg⁻¹ and 300 mg· kg⁻¹ YSE groups had particular suppressing effects on ruminal ammonia concentration, ammonia-N concentrations and protozoal populations. The effect of YSE on ruminal fermentation could be attributed to the selective inhibitory effect on rumen microbial species. High level (300 mg · kg⁻¹) YSE as feed additives resulted no negative impact on sheep in our tests.     

大豆LIM转录因子家族鉴定及盐胁迫响应分析

为探究大豆LIM转录因子家族基因在应答盐胁迫中的功能,利用生物信息学方法,根据大豆全基因组数据,共鉴定出21个LIM基因,命名为GmLIM01～GmLIM21,分析大豆LIM基因家族的染色体位置、蛋白质的理化性质、进化关系、保守基序以及对非生物逆境胁迫的响应。结果表明,这些GmLIM基因在大豆20条染色体中的14条染色体上分布不均,片段重复是大豆LIM家族扩大的主要原因。根据系统发育进化分析,大豆21个LIM基因可分为5个亚家族,与其他9个物种的LIM基因一起构建进化树,也可分为5个亚家族。大豆的21个GmLIM基因共包含10个基序,大部分LIM蛋白含有motif 1、motif 2和motif 4这3个保守基序。此外,顺式调控元件的分析表明,在GmLIM基因的启动子序列中,与光响应、生长发育、植物激素和非生物逆境胁迫应答相关的调控元件非常丰富。定量PCR结果表明,在盐胁迫处理后大豆叶和根中GmLIM07、GmLIM17基因的表达量分别在3和6 h达到峰值,后随着处理时间的加长表达量下降。综上GmLIM基因在大豆的盐胁迫应答过程中具有重要的调控作用。     

拟南芥 AtNHX5基因启动子的克隆和组织定位分析

为了探明拟南芥内膜反向转运体 AtNHX5基因启动子（proNHX5）功能及基因的组织表达模式,从基因组中克隆 AtNHX5基因开放阅读框（ORF）上游侧翼调控区1 807 bp序列,发现该启动子具有典型启动子一般特征,不仅具有TATA-box、CAAT-box等核心元件,还含有与光响应、激素响应、逆境诱导响应和分生组织表达调控元件。构建 AtNHX5基因启动子与GUS的融合表达载体,通过农杆菌花序浸染法转化野生型拟南芥获得转基因植株。利用组织染色法鉴定转基因拟南芥的GUS表达模式,发现在子叶、下胚轴和花中有显著的GUS酶活性,成熟叶片和根中只有局部检测到GUS表达,在未成熟果荚中只有在果荚顶端和基部存在GUS酶活性,说明 AtNHX5基因启动子与GUS的融合表达载体成功构建且正常启动GUS基因表达,同时也表明, AtNHX5基因主要在这些部位表达,且表达具有时空特异性。     

一个烟草糖基转移酶启动子在转基因烟草植株中的表达分析（摘要）

[目的]研究从烟草中克隆的1个受水杨酸和茉莉酸甲酯诱导表达的新的糖基转移酶基因（sm-Ngt）启动子部分缺失片段在烟草中的表达。[方法]以转sm-Ngt5个不同长度的5’端缺失的启动子与Gus基因融合植物表达载体的T1代转基因植株为材料,用茉莉酸甲酯（MeJA）和水杨酸（SA）处理16h后分别进行GUS组织化学染色和荧光定量法测定GUS酶活性,分析水杨酸和茉莉酸甲酯对sm-Ngt5个不同长度的5’端缺失的启动子表达的影响。[结果]在5个不同缺失片段启动子的转基因T1代生长30d的植株中,转-220～0bp片段的GUS染色最少,转-524～0bp及-468～0bp片段的染色最深。在没有MeJA和SA诱导处理时,-524～0bp和-468～0bp2个启动子片段启动的GUS活性最高,远高于-1150～0、-800～0、-220～0bp片段的活性,并且不是由于基因拷贝数而引起的（Southern杂交结果）;-800～0bp启动子片段启动的GUS活性受到MeJA和SA双重诱导,-1150～0bp启动子片段启动的GUS活性受到MeJA的诱导。[结论]在sm-Ngt启动子的-524～-220bp存在提高启动子活性调控元件,-1150～-524bp存在抑制启动子活性的序列,并且存在MeJA和SA双重诱导启动子活性调控元件。     

Accumulation and distribution of selenium in some vegetable crops grown in selenate-Se treated clay loam soil

A greenhouse experiment was conducted to study the accumulation of selenium by some vegetable crops commonly grown in the Indian Punjab. Eleven vegetable crops were raised in an alkaline clay loam soil treated with different levels of selenate-Se, i.e., 0, 1.25, 2.5 and 5.0 mg·kg⁻¹ soil. Dry matter yield of both edible and inedible portions of different vegetable crops decreased with increasing Se level in soil except potato (Solanum tuberosum), radish (Raphanus sativus) and cauliflower (Brassica oleracea var. botrytis) which recorded 10%–21% increase in inedible dry matter at 1.25 mg·kg⁻¹ Se soil. Application of 5 mg·kg⁻¹ selenate-Se soil resulted in complete mortality in the case of radish, turnip (Brassica rapa) and brinjal (Solanum melongena). Some vegetable crops including tomato (Lycopersicum esculentum), cauliflower and pea (Pisum sativum), though, survived the toxic effect at the highest concentration of Se yet did not bear any fruit. Potato and spinach (Spinacea oleracea) proved to be highly tolerant crops. Selenium concentration in the edible as well as inedible portions of all the vegetables increased with an increase in the level of applied Se. Selenium accumulation in the edible portion of vegetable crops in the no-Se control ranged from 2.2 to 4.9 mg·kg⁻¹ Se dry weight. At 1.25 mg·kg⁻¹ Se soil, the edible portion of radish accumulated the greatest concentration of Se (38 mg·kg⁻¹ Se dry weight) with that of onion (Allium cepa) bulb the lowest (9 mg·kg⁻¹ Se dry weight). Inedible portions of vegetables accumulated Se 2–5 times more than that absorbed by edible portions. Total Se uptake by edible portions of different vegetables was the greatest at 1.25 mg·kg⁻¹ Se soil, ranging from 7 to 485 μg·pot⁻¹. The results suggest that vegetable crops vary in their sensitivity to the presence of selenate-Se in soil. Vegetative portions were several times richer in Se than other parts of vegetable crops.     

不同品种紫花苜蓿氮代谢的Cd耐性评价及其鉴定指标筛选

为评估不同品种紫花苜蓿氮代谢对Cd的耐性并筛选可靠评价指标,以20个紫花苜蓿(Medicago sativa L.)品种为植物材料,以0、50 mg·kg~(-1)两个Cd处理浓度为土壤条件,开展温室盆栽试验。测定各品种的株高、生物量、全氮含量、硝态氮含量、脯氨酸含量、游离氨基酸含量、可溶性蛋白含量及氮代谢关键酶(硝酸还原酶、谷氨酰胺合成酶、谷氨酸合酶和谷氨酸脱氢酶)活性指标,计算Cd胁迫后各指标的变化率;分析各指标变化率之间的相关性,并利用主成分分析法和隶属函数法计算各品种的综合评价值(D),建立D值的回归模型并筛选Cd耐性的评价指标。结果显示,Cd胁迫后,11个指标在不同品种中的变化率各不相同,其中增幅最大的指标是品种"WL525HQ"中的硝态氮含量(122.24%),降幅最大的指标则是品种"WL525HQ"中的硝酸还原酶活性(-83.00%)。在株高、生物量、全氮含量、可溶性蛋白含量、脯氨酸含量、硝酸还原酶活性、谷氨酰胺合成酶活性和谷氨酸合酶活性8个指标之间,呈两两显著正相关关系(P0.05或P0.01)。20个紫花苜蓿品种中,"巨人"的D值最大(0.89),其氮代谢具有最强的耐Cd性,而"三得利"的D值最小(0.04),其氮代谢Cd耐性最弱。多元线性回归分析筛选出株高、谷氨酰胺合成酶、谷氨酸脱氢酶、硝态氮、硝酸还原酶和生物量6个单项指标作为评价紫花苜蓿氮代谢Cd耐性的主要指标,为紫花苜蓿Cd耐性品种的筛选与鉴定提供依据和数学模型。     

水稻植酸酶基因OsPHY2启动子的克隆及生物信息学分析

水稻种子萌发后,新生器官分化和生长所需磷素主要来自于植酸酶对种子中贮存植酸及其盐类的降解.针对水稻植酸酶基因OsPHY2在萌发种子中优势表达、但其转录调控机制尚不明确的现状,本研究克隆了该基因的启动子.利用生物信息学工具对该启动子中含有的顺式调控元件分析表明,该启动子中除含有RNA聚合酶结合的重要保守元件TATA盒和调...     

Influence of maize straw amendment on soil-borne diseases of winter wheat

A field experiment was conducted during the 2006–2007 wheat growing season at Baoding, Hebei Province, China, aiming at exploring the influence of different amendment rates of maize straw on winter wheat soil-borne diseases induced by Rhizoctonia cereali, Gaeumannomyces graminis and Bipolaris sorokiniana in field conditions. Wheat root vitality, ion infiltration, SOD activity, MDA content and microbial population of the tillage layer were measured. The results showed that the occurrence of three soil-borne diseases tested was significantly different under different amendment rates. During the greening stage and jointing stage, the disease indexes of three soil-borne diseases were reduced significantly by treatments at the maize straw amendment rates of 7500 kg · hm⁻² and 3750 kg · hm⁻². However, disease indexes of wheat common rot and sharp eyespot increased dramatically when the amendment rate increased to 15000 kg · hm⁻². At the amendment rate of 15000 kg · hm⁻², wheat root vigor and SOD activity decreased, and ion infiltration and cell membrane-lipid peroxidation level increased, respectively. In the meantime, higher amounts of bacteria and actinomycetes were recorded in the 7500 kg · hm⁻² amendment rate treatment, while a higher amount of fungi was recorded in the 15000 kg · hm⁻² amendment rate treatment.     

Bioinformatics analysis of tyrosinase-related protein 1 gene (<Emphasis Type="Italic">TYRP1</Emphasis>) from different species

As one member of the tyrosinase-related family directly involved in the production of melanin, TYRP1 is involved in not only melanogenesis but also prevention of melanocyte death, stabilizing tyrosinase and helping determine the shape of melanosomes, etc. Multi-species sequence comparisons showed that there were two evolutionally conserved non-coding regions (from −1306 to −733 and from −642 to −515 according to AL138753) upstream of translational initiation sites, representing putative regulatory regions subject to subsequent experimental tests. Coding sequence length variation and genetic diversity analysis showed that Felis catus, Homo sapiens and Canis familiaris had more genetic diversities than the other species for TYRP1, especially Felis catus that could be a better choice for studying the TYRP1-associated genetic basis underlying the color diversity. As a 75 kDa type-1 transmembrane glycoprotein, mature TYRP1 possesses about 17 kDa modifying components, whose function predominantly depends on the existing glycosyl-groups and the Cu components. In addition, the mutated amino acids within species and the highly conserved amino acids among species were listed in our paper.     

Study on the in vitro culture of cut plants in wheat haploid embryo induction by a wheat × maize cross

The wheat × maize system is one of the most effective ways to produce haploids in wheat. Whether and how it could be successfully applied in practical breeding mostly depends upon the efficiency of haploid embryo production. To perfect the protocols of haploid embryo induction, the efficiency of haploid embryo production between in vitro culture of cut plant and intact plant growth for hybrid spikes with two F₁ wheat hybrids and two maize varieties was compared. Effects of different cutting plant times and formulas of nutrient solutions for cut plant culture on haploid embryo formation were also studied. Results indicated that the embryo rate of in vitro culture was 3.29 times that of intact plant growth, with the figures of 31.6% vs 9.6%, respectively. The optimal time for cut plant culture was 24 h after pollination. Formulas of nutrient solutions significantly affected the efficiency of haploid embryo induction. With an embryo rate of 0–35.5%, adding calcium phosphate in the culture solution at 3 g·L⁻¹ could raise the caryopsis and embryo rates. According to this study, the best medium for cut plant culture was: 100 mg·L⁻¹ 2,4-D+ 40 g·L⁻¹ sucrose + 10 mg·L⁻¹ silver nitrate + 8 mL·L⁻¹ sulfurous acid + 3 g·L⁻¹ calcium phosphate, with which a caryopsis rate of 95% and an embryo rate of about 30% could be obtained. __________ Translated from Journal of Triticeae Crops, 2008, 28(1): 1–5 [译自: 麦类作物学报]     

Influences of nitrogen fertilizer application rates on radish yield, nutrition quality, and nitrogen recovery efficiency

Radishes (Raphanus sativus L.) were grown in plastic pots in a screenhouse to investigate the influences of nitrogen fertilizer application rates (NFAR) on yield, nitrate content, nitrate reductase activity (NR), nutrition quality, and nitrogen recovery efficiency (NRE) at commercial mature stage. Five N-rate treatments, 0.644, 0.819, 0.995, 1.170, and 1.346 g·pot⁻¹, were set up in the screenhouse pot experiments, and nitrogen fertilizer (unlabeled N and ¹⁵N-labeled fertilizer) was applied as basal dressing and topdressing, respectively. The results indicated that the fresh and dry weight yields of radish increased with the increase of NFAR at the range of 0.099 to 0.180 g N·kg⁻¹ soil, decreased at 0.207 g N·kg⁻¹ soil, and accordingly there was a significant quadratic relationship between the fresh and dry weight yields of radish and the NFAR. At the high addition of urea-N fertilizer, the nitrate content accumulated in the fleshy roots and leaves due to the decline in NR activity. From 0.644 to 0.819 g N·pot⁻¹ NR increased most rapidly, the highest NR activity occurred at 0.819 g N·pot⁻¹, and the lowest NR activity happened at 1.346 g N·pot⁻¹. Soluble sugar and ascorbic acid initially increased to the highest value and then decreased, and, contrarily, crude fiber rapidly decreased with the increase of NFAR. Total N uptake (TNU), N derived from fertilizer (N_dff), and N derived from soil (N_dfs) in radish increased, except that N_dfs relatively and slightly decreased at the rate of 0.207 g N·kg⁻¹soil. The ratio of Ndff to TNU increased, but the ratio of Ndfs to TNU as well as NRE of N fertilizer decreased with the increase of NFAR. Therefore, the appropriate NFAR should be preferably recommended for improving the yields and nutrition qualities of radish and NRE of N fertilizer. These authors contributed equally to this work     

甘蓝型油菜种子特异表达油体蛋白启动子P_(BnOA03)的克隆及功能分析

为了改良油菜种子的性状,常常需要研究种子特异表达目的基因的情况。在分析油菜油体蛋白表达模式的基础上,选择种子特异高表达的油体蛋白,根据甘蓝型油菜基因组序列设计引物,利用PCR技术从基因组中克隆到一段715bp的启动子序列。启动子顺式元件分析表明,这一克隆序列具有CAAT-box和TATA-box启动子基本元件,另外含有ABA响应元件等多种顺式元件。进一步利用GUS报告基因在拟南芥中对该启动子的功能进行鉴定,结果表明,在转基因株系的幼苗、根、茎、叶、花和种子发育前期及成熟种子中没有检测到GUS基因明显表达,在种子发育的中后期检测到GUS基因的表达逐渐增强。说明这个启动子的表达调控具有一定的种子特异性。     

十字花科植物PIN3基因调控元件及表达分析

为探明十字花科不同物种PIN3基因的表达调控差异,从拟南芥(Arabidopsis thaliala)、红花荠菜(Capsella rubella)、白菜(Brassica rapa)、油菜(Brassica napus)和甘蓝(Brassica oleracea)等17个已完成基因组测序的十字花科物种基因组数据库中鉴定出18个PIN3同源基因。对这18个同源基因编码序列上游1 500bp启动子区域顺式作用元件进行预测及比对分析,结果发现,光响应、多种激素响应及胁迫响应等相关元件呈现出较大差异,其中生长素以及向地性响应相关元件具有较高的保守性。克隆普通荠菜PIN3同源基因(CbPIN3)并构建普通荠菜PIN3基因启动子的GUS报告系统CbPIN3pro::GUS转化拟南芥。与拟南芥基因表达数据库比较,荠菜启动子表达与拟南芥PIN3具有基本一致的组织表达模式,但也存在一定的组织差异,说明植株形态建成中生长素极性转运调控的差异。拟南芥和荠菜PIN3的表达差异是十字花科物种适应性进化模式的缩影。     

Effects of dietary squid viscera meal on growth and cadmium accumulation in tissues of large yellow croaker, Pseudosciaena crocea R.

Cadmium (Cd) is a toxic environmental pollutant with a long biological half-life and can produce both hepatic and renal injuries in mammals and fish. Squid viscera meal (SVM), an effective attractant for aquatic animals, is widely used as an ingredient in aquafeeds. However, SVM is rich in Cd and its complexes. A study was conducted to evaluate the effects of dietary SVM on the growth and Cd deposition in the tissues of large yellow croaker, Pseudosciaena crocea R. Three practical diets were formulated to contain a 0, 50 and 100 g · kg⁻¹ SVM diet, correspondingly containing a 0.21, 7.26 and 12.08 mg Cd · kg⁻¹ diet. Each diet was randomly assigned to triplicate groups of 100 juveniles of large yellow croaker (mean initial weight, 9.75 ± 0.35 g) in floating sea cages (1.0 m × 1.0 m × 1.5 m). Fish were fed twice daily (05:00 and 17:00) to satiation for 8 weeks. The results showed that there were no significant differences in fish survival among the three dietary treatments, but significant higher specific growth rates (SGR) were observed in the fish fed diets with 50 or 100 g · kg⁻¹ SVM diet compared to the control group (P < 0.05). The cadmium concentrations in fish tissues (muscle, liver, kidney and gill) were significantly influenced by the dietary SVM. The cadmium concentrations in all tissues significantly increased with increasing dietary Cd levels (P < 0.05). In all the dietary treatments, the highest Cd level was always observed in the kidney, followed by the liver and the gill. Fish fed diets with 50 and 100 g · kg⁻¹ SVM had significantly higher Cd accumulations in the kidney (2.65, 4.44 mg · kg⁻¹), liver (0.58, 0.93 mg · kg⁻¹) and gill (0.35, 0.53 mg · kg⁻¹) compared with the control group (0.42, 0.26 and 0.12 mg · kg⁻¹, respectively). The Cd level in fish muscle, however, was undetectable in all treatments. Therefore, based on these results, accumulation of Cd in edible tissue (muscle) of farmed large yellow croaker is not a food safety issue. However, long-term feeding of diets with SVM may result in accumulation of Cd in the kidney, liver and gills of fish.     

魔芋GDP甘露糖焦磷酸化酶基因及其启动子的克隆与分析

以花魔芋和白魔芋为材料,通过魔芋5个转录组高通量测序数据库,与拟南芥进行tBlastn比对、拼接,得到魔芋中的GDP-甘露糖焦磷酸化酶基因GMP的推定全长,设计基因特异性引物,克隆得到魔芋GMP基因的部分cDNA序列,长度为1 233 bp,其中基因编码区长度为1 086 bp,编码的氨基酸为361个.在此基础上运用FNPI-PCR技术得到了魔芋GDP-甘露糖焦磷酸化酶基因GMP上游启动子2 116 bp,经生物信息学分析,该序列具有典型的TATA-box、 CAAT-box及与胁迫相关的元件和光应答元件.