大白菜种子蛋白分离的高效液相色谱法的研究

该文首次利用高效液相色谱法时大白菜种子的水溶性蛋白质进行了分析,确定了适宜的色谱分析条件:选用径向加压型Nova-Pak C_(18)反相色谱柱(规格为8mm (?).d.×10cm length,填充颗粒为球形,直径4μm,Waters Co.)以甲醇水溶液进行洗脱,种子中各蛋白组分得到了比较好的分离。洗脱时采用混合梯度洗脱方式:0-20分钟,从9%甲醇水溶液(A)到35%甲醇 0.02%三氧乙酸(B)的线性梯度洗脱;20-45分钟,以B洗脱液进行等强度洗脱.流速均为0.8ml/min。洗脱后的各组分用紫外光/可见光检测器在280nm处进行监测。实验结果还表明:不同提取液对色谱分离的结果有一定的影响,用0.1%巯基乙醇水溶液提取时,色谱峰的分离效果较好,主要组分的色谱峰比较显著,因而比较适于大白菜种子的分析。     

大白菜的类胡萝卜素种类鉴定

为建立大白菜类胡萝卜素鉴定方法,利用反相高效液相色谱（RP-HPLC）,根据标样和二极管阵列检测器的检测结果,对不同类型大白菜材料的类胡萝卜素种类进行鉴定。结果表明：利用C30柱从大白菜中共分离出48类胡萝卜素,根据保留时间和吸收谱特征鉴定出其中的30种成分,链孢红素、玉米黄质、六氢番茄红素、ζ-胡萝卜素、γ-胡萝卜素和紫黄质均为首次在大白菜中报道。橙色大白菜中的类胡萝卜素种类最多,与其他大白菜存在很大差异。说明该鉴定方法适用于大白菜的类胡萝卜素种类研究。     

含氟杀虫剂类农药的残留检测分析研究进展

由于氟原子所具有的特殊功效,近几十年来含氟类农药发展迅速。含氟杀虫剂主要包括拟除虫菊酯和苯甲酰脲中的含氟品种,以及氟虫腈、溴虫腈、茚虫威等杂环类化合物。本研究对含氟杀虫剂类农药的残留分析方法进行了综述,包括样品提取技术、净化技术及检测技术。样品净化技术包括固相萃取和液-液萃取,检测技术主要有气象色谱法、高效液相色谱法、气相色谱-质谱联用(GC-MS)和高效液相色谱-质谱联用(HPLC-MS)。最后从动物性食品安全的角度提出建议。     

烟草中茄尼醇检测方法研究进展

为了给烟草中茄尼醇的进一步研究、开发与应用提供参考,笔者对目前应用于烟草中茄尼醇分析的主要检测方法作了较详细评述。方法包括高效液相色谱法、气相色谱法、液相色谱-质谱联用法、气相色谱-质谱联用法、薄层色谱法、近红外漫反射技术、魔角旋转核磁共振法、库仑滴定法和间接碘量法,并对目前烟草中茄尼醇的检测方法的缺点进行了分析,对未来的发展方向进行了展望。     

不同种质及生长年限关黄柏中生物碱含量变化规律的研究

目的：采用反相高效液相色谱法对不同种质（北京、东北）,不同生长年限关黄柏中有效成分（小檗碱、巴马汀、药根碱）含量的变化规律进行研究。为科学地确定关黄柏采收期,保护关黄柏资源提供依据。方法：高效液相色谱法,色谱柱:Alltima HP C18（250×4.6mm,5μm）,流动相：乙腈-0.1%磷酸水溶液（40:60,含0.1%的十二烷基磺酸钠）,流速：1.0ml/min;柱温：25℃,检测波长：345nm。结果：北京种质2年生以上、东北种质（吉林通化）9年生以上的关黄柏中小檗碱含量可达到中国药典规定的含量要求。结论：不同种质,不同生长年限的关黄柏生物碱含量具有差异性,且变化趋势不同。     

氟喹诺酮类兽药残留检测方法研究进展

综述了动物性食品中氟喹诺酮类药物残留检测的微生物法、液相色谱法、高效液相色谱法、高效毛细管电泳分析法、液相色谱/串联质谱法和酶联免疫法的研究进展,为进一步开展药物残留检测的研究和监测提供借鉴。     

草甘膦农药残留检测方法研究进展

草甘膦农药因具有低毒、高效的特点,目前已成为全球生产及使用量最大的除草剂之一,但其潜在毒性近年来也引起了人们的广泛关注,并且对于草甘膦残留的检测研究也备受重视。对草甘膦残留的检测方法及研究进展进行综述,主要探讨了化学分析法、酶联免疫吸附测定法、分光光度法、高效液相色谱法、高效液相色谱-质谱联用法、气相色谱法、气相色谱-质谱联用法、电化学方法在草甘膦残留检测中的应用,为草甘膦残留检测深入研究提供参考。     

AFLP分子标记鉴别大白菜品种

本试验采用AFLP技术,研究了90份来自7个不同栽培地区的大白菜品种材料。共筛选了20对引物,不同引物组合检测多态性谱带的能力有很大的差异,多态性谱带的数量从9条到32条不等。其中E—ACA／M—CTG是大白菜品种十分高效的引物组合,共产生7l条清晰的扩增带,其中有32条多态性谱带,多态性谱带的百分率为45．7％。通过该引物组合,能将90个品种全部区分开来。同时应用该引物组合检测2个大白菜杂交品种(北京新2号,京夏王)各10株,其中有1株北京新2号的谱带异常,其余同一品种不同单株的带型完全一致。表明AFLP标记用于研究品种指纹图谱,并鉴别品种是完全可行的。     

浅谈常规蔬菜农残检测技术

常规蔬菜的农残检测技术,是在食品安全问题中最基础的保障和关键技术。通过对常规蔬菜农残检测步骤及运用的多种检测技术进行分析,对检测技术中的气相色谱法、气相色谱-质谱联用法、高效液相色谱法、液相色谱-质谱联用法和酶抑制法等检测技术进行论述,研究各种技术在检测蔬菜农残中的应用。     

生姜中农药残留检测方法

食品农药残留问题成为人们普遍关注的社会热点问题,生姜中农药检测方法主要有气相色谱法、气相色谱-质谱法、液相色谱-电喷雾串联质谱法和超高效液相色谱-串联质谱法等。对这些检测方法进行系统总结,以期为生姜农药进一步检测提供技术选择参考。     

Studies on glucosinolates in clubroot resistant selections and susceptible commercial cultivars of cabbages

Summary Heads of 59 commercial cabbage cultivars, all susceptible to clubroot disease, and of 86 individual clubroot resistant cabbage plants of various breeding selections were analysed for their composition in glucosinolates, determined by quantifying their hydrolytic breakdown products, thiocyanate, goitrin, and volatile isothiocyanates. The mean thiocynate ion content was significantly lower in the breeding selections (120 g/g dry weight) than in the commercial cultivars (204 g/g). In contrast, the mean goitrin content was significantly higher in the breeding selections (193 g/g) than in the commercial cultivars (35 g/g). Similar to goitrin, the range of volatile isothiocyanates and total glucosinolate were higher in the breeding selections, but the mean contents of each were not statistically different between selections and cultivars. Fourteen cultivars and four selections were found to be free of goitrin; three cultivars, but no breeding selection were free of volatile isothiocyanates. The breeding selections will provide germplasm for breeding new clubroot resistant and low glucosinolate cultivars.Contribution No. J-965.     

盐胁迫对大白菜幼苗生理生化特性的影响

为了探讨大白菜耐盐机理、筛选耐盐性鉴定指标,建立苗期耐盐性鉴定方法,选用了６个具有不同耐盐性的大白菜材料,用Hoagland营养液水培方法,研究了不同浓度NaCl胁迫处理下大白菜盐害指数、脯氨酸含量、可溶性糖含量、游离氨基酸含量和丙二醛（MDA）含量的变化规律。结果发现,随着NaCl处理浓度的增加以及胁迫时间的延长,大白菜幼苗叶片中的MDA含量、脯氨酸含量、可溶性糖含量以及游离氨基酸含量均呈上升趋势。不同大白菜品种的盐害指数表现出不同程度的增加。盐害指数和MDA含量在不同品种间表现了明显差异,能较好地反映大白菜各品种苗期的耐盐性,可作为大白菜耐盐性的筛选指标。     

成都平原主栽大白菜品种根肿病抗性的鉴定

为明确成都平原地区主栽大白菜品种对根肿病的抗性情况,于2018年分别在成都市新都区、彭州市和崇州市3个试验田块对45个大白菜主栽品种采用田间抗鉴圃自然感染诱发法进行根肿病抗性鉴定,通过调查根部根肿病发病程度评价各品种的抗性。结果表明,在崇州自然病圃中表现为免疫的品种有4个,占总品种的8.9%;其余品种均表现为感病或高感,占比91.1%。彭州的结果显示,在所测定的45个品种中,免疫品种有6个;耐病品种有11个;剩余28个品种为感病或高感品种。而在新都发病田块中,只有2个品种表现为对根肿病免疫,16个品种表现为耐病,27个品种表现为感病或高感。部分品种在不同地区抗性结果表现不一致,试验鉴定到2个对根肿病免疫的优异品种,可为选育和应用抗性品种提供借鉴。     

Genetic relationships of Brassica vegetables determined using database derived simple sequence repeats

Sequence databases were screened to identify simple sequence repeats (SSRs) in Brassica oleracea sequences. A total of 512 B. oleracea DNA sequences were screened and 43 potential SSRs were identified. Thirty-six primer pairs were designed to amplify target sequences. Of the 36 primer pairs, six failed to amplify fragments of expected sizes, and 17 primer pairs failed to generate polymorphisms. Thirteen SSRs were used to assess genetic similarity between 54 B. oleracea cultivars, belonging to 3 variteal groups (cabbage, cauliflower, and broccoli). Pairwise genetic similarities were calculated for cultivars, and a dendrogram of relationships was produced. All cabbage cultivars were distinguished from each other and clustered in two separate groups. Five cauliflower cultivars could not be distinguished with SSR markers used in the study. Three broccoli cultivars clustered with cauliflower cultivars, and two cauliflower cultivars grouped with broccoli cultivars. The varietal group with the narrowest genetic variation in the study was cauliflower (B. oleracea var. botrytis) followed by broccoli (B. oleracea var. italica) and cabbage (B. oleracea var. capitata) groups. Polymorphism information content (PIC) values and number of alleles produced per marker ranged between 0.25 to 0.86 and 1 to 8, respectively, for database derived SSR markers.     

Assessment of DNA markers for seed contamination testing and selection of disease resistance in cabbage

Cabbage (Brassica oleracea L. var. capitata) is an important vegetable worldwide. Most Japanese commercial cultivars of cabbage use an F₁ hybrid seed production system. The purity of F₁ hybrid seeds is important and the assessment of purity based on DNA markers can be highly accurate. In addition, selection of agronomically important traits such as disease resistance based on DNA markers is useful for breeding of cabbage. The aim of this study is to demonstrate the effectiveness of DNA marker-assisted selection in cabbage. In this study we distinguished the parental S haplotypes in 35 F₁ hybrid cultivars by combining several linked DNA markers. Thirty-one highly polymorphic simple sequence repeats (SSR) markers were screened from 175 reported SSR markers, which are useful for assessment of the purity of F₁ hybrid seeds. We examined the relationship between the DNA marker based genotype and the phenotype by an inoculation test of clubroot disease. A co-dominant PCR–RFLP marker was developed for selection of Fusarium yellows resistance and the genotypes using this marker were consistent with inoculation test in all tested samples.     

利用RAPD技术鉴定大白菜主栽品种及品种间遗传多样性分析

采用随机引物扩增多态ＤＮＡ（ＲＡＰＤ）技术分析了２１个大白菜主栽品种。用１３个引物共扩增出８７个清晰可重复的ＤＮＡ片段,其中３９条带具有多态性,多态性条带的频率为４４．８％。ＯＰＥ０１是多态性频率最高的引物,它可区分１５个大白菜品种,再与引物ＯＰＨ０３和ＯＰＨ１２配合,即可将２１个大白菜品种区别开。     

大白菜一代杂种及双亲幼苗的等电聚焦电泳过氧化物同工酶分析

等电聚焦电泳是近些年发展起来的分离蛋白质的有效生化技术．操作简便，分辨率高，电泳条带多而且细，信息量大，利于进行深入细致的研究。该文利用等电聚焦丙烯酰胺电泳方法分析了6个大白菜一代杂种及双亲幼苗的过氧化物同工酶电泳图谱，共分离出十几条酶带，F1杂种与亲本间同工酶图谱均表现出明现差异，特别是A鉴别区内的酶带差异最明显，大多为互补型，是主要鉴别区。进一步研究结果表明，6个大白菜品种均可利用过氧化物同工酶鉴别由自交引起的假杂种。利用这项技术大量开展了这6个品种的商品种子纯度检测工作，同时在田间进行了验证性测定，二者结果基本相符。     

检测方法对甘蓝和大白菜品种RAPD标记鉴别多态性的影响

分析了琼脂糖凝胶电泳和聚丙烯酰胺凝胶电泳2种检测方法对甘蓝和大白菜品种RAPD标记鉴定多态性水平的影响.用12个随机引物扩增了14个甘蓝品种,琼脂糖凝胶电泳检测的结果为平均每个引物产生4.5条扩增带,其中1.5条具有多态性,多态性频率为33.3％.同时对产生条带最多的引物(S149)所扩增的产物进行聚丙烯酰胺凝胶电泳检测,结果检测到了30条清晰可见的扩增带,其中19条具有多态性,为前者的6倍,可以完全鉴别14个甘蓝品种.用5个随机引物扩增了23个大白菜品种,用两种检测方法同时检测了扩增结果.结果表明,琼脂糖凝胶电泳检测的平均多态频率为30％,聚丙烯酰胺凝胶电泳检测平均多态频率为61％,后者为前者的两倍.因此,在使用RAPD标记进行甘蓝和大白菜品种鉴别时,聚丙烯酰胺凝胶电泳检测方法能大大提高检测效率.     

An S-allele survey of cabbage (Brassica oleracea var. Capitata)

Summary A total of 31 S-alleles was found in a survey of 197 cabbage plants representing 11 cultivars of diverse type. Most of these S-alleles also occurred in either kale or Brussels sprouts, but five of them have not been found previously and apparently occur only in cabbage. A more detailed study of five cultivars of spring cabbage showed only 12 S-alleles in all, with 6–10 S-alleles in four older cultivars and only 3 S-alleles in the newer more highly selected cultivar. S2 was by far the commonest S-allele, as it is in B. oleracea as a whole. The highly recessive alleles S5 and S15 were not particularly common in cabbage and this may partly explain why the sib problem in F₁ hybrids is apparently less in cabbage than in Brussels sprouts. Three cases were found in which an S-allele was completely recessive in both the stigma and the pollen. The problems for the breeder created by this rather unusual situation are discussed.