中间锦鸡儿FAD2基因拷贝数检测及组织表达谱分析

【目的】探索中间锦鸡儿基因组中FAD2基因的拷贝数,分析不同拷贝的表达模式,以期揭示不同拷贝在植物体内的功能分工。【方法】根据中间锦鸡儿(Caragana intermedia)FAD2基因的保守序列设计1对引物SF和SR,利用该引物对PCR扩增制备114 bp的Southern检测探针,探针进行地高辛标记后,采用罗氏Southern杂交试剂盒进行Southern检测。根据中间锦鸡儿3个FAD2基因各自的特异差异序列,分别设计定量PCR引物序列,以actin基因为内参,对中间锦鸡儿的根、茎、叶及不同发育时期(发芽15,25,35 d)种子的cDNA进行定量PCR分析。【结果】中间锦鸡儿FAD2基因至少有4个拷贝,且不同拷贝的表达模式不同。FAD2-2A基因在根和发育中期、后期的种子中低水平表达,在幼嫩的茎、叶以及发育早期的种子中高水平表达;FAD2-1A基因在根中的表达量最低,在种子发育早期、中期大量表达,在幼嫩叶子中表达水平也较高;FAD2-1B基因仅在种子发育中期大量表达,在其他组织及种子其他发育阶段的表达量均保持在本底水平。在不同组织以及种子的不同发育时期,FAD2-1A相对表达量的变化幅度最大,达到了近100倍,FAD2-1B次之,FAD2-2A最小。【结论】结合序列比对分析推测:FAD2-2A基因可能主要负责合成膜脂中的亚油酸,FAD2-1A主要负责种子及叶子贮脂中亚油酸的合成,FAD2-1B负责种子贮脂中油酸的去饱和作用。     

光照和温度对中型滇丁香种子萌发的影响

采用室内发芽试验法研究了不同光照和温度条件对中型滇丁香种子萌发的影响。结果表明,中型滇丁香种子的萌发需在有光照的条件下进行,在有光照状况下中型滇丁香种子萌发的最适温度为25℃。在此温度条件下,其种子12～14天开始萌发,2～3周萌发完全,种子发芽率可达98%。以25℃为界,温度的升高或降低均会降低其种子的发芽率。     

高寒沙地2种锦鸡儿的根系分布

中间锦鸡儿和柠条锦鸡儿是共和盆地沙丘造林的主要树种。通过剖面法比较了2种锦鸡儿10年生人工林的不同级别的根系分布特征。结果表明:2种锦鸡儿均为深根系植物,深度达到1 m。中间锦鸡儿的根系生物量主要分布在0~60 cm。其中,粗根、中根和细根分别主要分布在0~30、0~60 cm和0~60 cm。然而,它的3个不同径级根系的长度分别主要分布在0~30、0~50 cm和0~60 cm。柠条锦鸡儿的根系生物量主要分布在0~90 cm。其中,粗根、中根和细根分别主要分布在0~60、0~60 cm和0~90 cm。然而,它的3个不同径级根系的长度分别主要分布在0~60、0~60 cm和0~70 cm。因此,中间锦鸡儿的根系相对较浅,适宜吸收降雨补充的浅层和中层土壤水分。柠条锦鸡儿的根系向更深的土壤中生长,吸收深层土壤水分。这反映了2种锦鸡儿对高寒沙地半干旱气候的适应差异。     

The effect of geographic location, breed, and pituitary dysfunction on seasonal adrenocorticotropin and α-melanocyte-stimulating hormone plasma concentrations in horses

Background: Plasma α‐melanocyte‐stimulating hormone (α‐MSH) and adrenocorticotropin (ACTH) concentrations in horses vary with season, confounding diagnostic testing for pituitary pars intermedia dysfunction (PPID). Hypothesis: The goals of this study were to determine whether seasonal variation in plasma α‐MSH and ACTH concentrations in horses is influenced by geographic location, breed, or PPID. Animals: Healthy light breed horses residing in Florida, Massachusetts, and Finland (n = 12 per group); healthy Morgan horses (n = 13); healthy ponies (n = 9) and horses with PPID (n = 8). Methods: Monthly plasma α‐MSH and ACTH concentrations were measured by radioimmunoassay. Nonlinear regression analysis was used to estimate the time of peak hormone concentrations. Mean hormone concentrations in fall and nonfall months were compared. Results: The fall peak plasma α‐MSH concentration occurred earlier in horses residing at more northern locations. Mean seasonal α‐MSH concentrations were similar in all healthy groups at all locations, but in the fall, plasma ACTH concentrations were higher in horses living in more southern locations. Plasma ACTH but not α‐MSH concentrations were higher in Morgan horses compared with light breed horses from the same location. Hormone concentrations of ponies did not differ from those of horses during either season. Concentrations of both hormones were high in the fall compared with the spring in horses with PPID. Conclusions and Clinical Importance: These findings suggest geographic location of residence and breed may affect the onset, amplitude, or both of the seasonal peak of pars intermedia (PI) hormones and should be considered when performing diagnostic testing for PPID. Horses with PPID maintain seasonal regulation of PI hormone output.     

Diagnostic orientation values for ACTH and other parameters for clinically healthy donkeys and mules (insulin,triglycerides, glucose,fructosamines, and ɣ-GT)

Pituitary pars intermedia dysfunction is the most prevalent endocrine disease in horses. Although donkeys and mules may also be affected, only a few data have been published. Reference values for diagnostic parameters, such as adrenocorticotropic hormone (ACTH), are especially scarce or even lacking. Therefore, in the present study, available data from the literature have been verified and completed to facilitate a reliable diagnosis. Clinical inspections and haematological and biochemical examinations were carried out four times in a three-month interval (February to November) in 44 donkeys and 31 mules. Data from clinically healthy animals were used as an orientation. Plasma ACTH concentrations showed seasonal changes in both animal groups. However, it was generally higher in donkeys than mules. Although blood glucose (EDTA plasma) showed no difference between groups, serum insulin concentrations were consistently higher in donkeys. Serum fructosamine levels were slightly higher in mules, whereas, in some cases, serum triglyceride levels were considerably higher in donkeys. Serum gamma-glutamyltransferase showed a striking peak in mules in August, whereas the remaining gamma-glutamyltransferase values were lower compared to donkeys. By comparing donkeys and mules, the present work reveals differences in various blood parameters which should be considered for diagnoses and future studies.     

中麻黄生境及栽培因子研究

通过对河西地区中麻黄资源生境调查和栽培学因子及生长规律的试验研究表明:在降水量较大的河西东部,天然中麻黄分布群落较多;随着降雨量递降,中麻黄种群数量和面积递减,生长势也渐弱。中麻黄在轻、中度盐渍化土壤中生长良好,而在重度盐碱地上稀见分布。中麻黄人工种植中,当土壤含盐量在0.2 % 时,对麻黄幼苗的影响小;当土壤含盐量达0.82 % 时,出苗率为41 %,盐害死亡率达48.8 %;当土壤含盐量达0.76 % 时对当年生麻黄苗产生影响。中麻黄虽是强旱生耐瘠薄的植物,但通过合理施肥,适量灌溉等栽培技术能促进其生长发育,只需耕作农田年灌水量的1/2～1/6以及有机肥和速效氮、磷肥为主的适量施肥,就可大大提高单位面积上中麻黄的产量,提高麻黄碱含量。     

中间偃麦草、长穗偃麦草及其杂种F1代同工酶研究

通过对过氧化物酶(POD)、酯酶(EST)、超氧化物歧化酶(SOD)和多酚氧化酶(PPO)同工酶谱的分析,探讨中间偃麦草(Elytrigia elongata(Host)Nevski)、长穗偃麦草(E.intermedia(Host)Nevski)及其杂种F₁代的酶谱特征和亲缘关系。结果表明:亲本同工酶酶带数目、迁移率、着色程度等差异不大,二者亲缘关系较近。与亲本相比,杂种F₁代存在POD、SOD、PPO酶带丢失现象,但正、反交植株中均产生了一些亲本没有的新生带。杂种F₁代EST酶谱与亲本基本相同,仅着色程度略有差异。杂种F₁代植株的POD、PPO酶谱特征有偏向中间偃麦草的倾向,而SOD酶谱特征倾向于长穗偃麦草。杂种F₁代同工酶具有多态性,可作为遗传标记用于杂种鉴定和后代株系目标性状的选择。     

中间锦鸡儿fad72基因克隆与序列分析

油酸去饱和酶FAD2（fatty acid desaturase 2）在油酸中引第二个双键生成亚油酸,是负责植物体内产生多不饱和脂肪酸的第一步关键酶。本研究采用RT—PCR和RACE（rapid amplification of cDNA ends）技术,以中间锦鸡儿未成熟种子为材料,克隆了三个fad2基因（分别命名为fad2-2A,fad2-1A和foa2-1B）。将这三个基因与汪阳东从中间锦鸡儿枝叶中克隆的fad2—2B（CaFAD2基因,GenBank登录号AY957394）一起进行了序列比对和分析。序列同源性比较结果表明,fad2—1A与fad2—1B同源性很高,fad2—1A编码的前283个氨基酸与fad2-1B的同源性高达98．9％,fad2—2B与faa2—1A的氨基酸序列同源性最低,只有64．7％。这四个基因的成功克隆,为进一步研究基因分工、调控方式打下了基础,为实现对锦鸡儿属植物脂肪酸成分直接、精确调控提供了保证,也是从一个物种中克隆四个fad2基因的第一次报道。     

中间锦鸡儿硬枝扦插试验

以中间锦鸡儿为材料进行了硬枝扦插试验,在珍珠岩和粗沙2种不同的基质上进行扦插,分别采用不同浓度的吲哚丁酸(IBA)、奈乙酸(NAA)、ABT1#、ABT2#生根粉进行处理。试验表明:粗沙硬枝扦插较为理想,平均生根率较珍珠岩提高14.1%;不同激素种类对中间锦鸡儿硬枝插穗生根情况影响极显著,珍珠岩基质上扦插,以ABT1#100 ppm和200 ppm生根效果最佳;粗沙基质上扦插,处理浓度以IBA 1000 ppm、ABT1#500 ppm、ABT2#500 ppm对中间锦鸡儿硬枝扦插生根效果最佳。     

中间锦鸡儿fad2基因克隆与序列分析

油酸去饱和酶FAD2(fatty acid desaturase 2)在油酸中引第二个双键生成亚油酸,是负责植物体内产生多不饱和脂肪酸的第一步关键酶.本研究采用RT-PCR和RACE(rapid amplification of cDNA ends)技术,以中间锦鸡儿未成熟种子为材料,克隆了三个fad2基因(分别命名为fad2-2A、fad2-1A和fad2-1B).将这三个基因与汪阳东从中间锦鸡儿枝叶中克隆的fad2-2B(CaFAD2基因,GenBank登录号AY957394)一起进行了序列比对和分析.序列同源性比较结果表明,fad2-1A与fad2-1B同源性很高,fad2-1A编码的前283个氨基酸与fad2-1B的同源性高达98.9%,fad2-2B与fad2-1A的氨基酸序列同源性最低,只有64.7%.这四个基因的成功克隆,为进一步研究基因分工、调控方式打下了基础,为实现对锦鸡儿属植物脂肪酸成分直接、精确调控提供了保证,也是从一个物种中克隆四个fad2基因的第一次报道.