油菜硫苷组分含量及抑菌活性研究

对油菜不同器官硫苷的组分与含量进行分析,结果表明根系和茎中的硫苷总量显著高于叶片,根系中的硫苷以4甲-氧基-3吲-哚甲基硫苷和1-甲氧基-3吲-哚甲基硫苷为主,茎和叶中以2羟-基-3丁-烯基硫苷和苯乙基硫苷为主。油菜根系中的硫苷组分从冬前期到终花期基本保持稳定,硫苷各组分的含量在不同生育时期的变化基本一致,终花期含量最高,成熟期含量最低。19个油菜品种根系中硫苷组分与含量存在较大差异;以草莓灰霉菌为靶标物,研究油菜根系的挥发性水解产物和水溶性水解产物对土传病原真菌的抑制作用,发现不同油菜品种根系水解产物的抑菌活性与其硫苷组分与含量有关,草莓灰霉菌对丁烯基ITC非常敏感,吲哚类ITCs对草莓灰霉生长的抑制作用要低于芳香族ITCs。     

油菜油分、蛋白质和硫苷含量相关性分析及QTL 定位

为定位与油分、蛋白质和硫苷含量等品质性状相关的数量性状位点(QTL), 以2个含油量较高的甘蓝型油菜(Brassica napus)品系8908B和R1为研究材料, 配置正反交组合。在正反交F₂代群体中, 含油量和蛋白质含量都存在极显著的负相关, 相关系数分别为-0.68和-0.81, 含油量和硫苷含量相关性不显著; 蛋白质含量和硫苷含量在正交群体中相关性不显著, 但在反交群体中存在显著负相关(相关系数r =-0.45)。利用正交F₂代群体中的118个单株, 构建了包含121个标记的遗传连锁图谱, 图谱长1 298.7 cM, 有21个连锁群(LGs)。采用复合区间作图法, 在连锁图上定位了2个与含油量有关的QTL, 分别位于LG8和LG10, 其贡献率分别为4.8%和13.7%, 增效基因都来源于R1; 定位了2个与蛋白质含量有关的QTL: pro1 和 pro2, 分别位于LG1和LG3, 其贡献率分别为15.2%和14.1%, 位点pro1由8908B提供增效基因, pro2则由R1提供增效基因; 定位了4个与硫苷含量有关的QTL, 其中LG20上有2个, LG4和LG8上各1个, 它们的贡献率在1.9%-25.4%之间, 除LG20上glu1的增效基因来自R1外, 其余3个QTL位点均由8908B提供增效基因。     

油菜油分、蛋白质和硫苷含量相关性分析及QTL定位

为定位与油分、蛋白质和硫苷含量等品质性状相关的数量性状位点（QTL）,以2个含油量较高的甘蓝型油菜（Brassica napus）品系8908B和R1为研究材料,配置正反交组合。在正反交F2代群体中,含油量和蛋白质含量都存在极显著的负相关,相关系数分别为-0．68和-0．81,含油量和硫苷含量相关性不显著：蛋白质含量和硫苷含量在正交群体中相关性不显著,但在反交群体中存在显著负相关（相关系数r=-0．45）。利用正交F2代群体中的118个单株,构建了包含121个标记的遗传连锁图谱,图谱长1298．7cM,有21个连锁群（LGs）。采用复合区间作图法,在连锁图上定位了2个与含油量有关的QTL,分别位于LG8和LG10,其贡献率分别为4．8％和13．7％,增效基因都来源于R1;定位了2个与蛋白质含量有关的QTL：pr01和pr02,分别位于LG1和LG3,其贡献率分别为15．2％和14．1％,位点pr07由8908B提供增效基因,pro2则由R1提供增效基因：定位了4个与硫苷含量有关的QTL,其中LG20上有2个,LG4和LG8上各1个,它们的贡献率在1．9％-25．4％之间,除LG20上glu7的增效基因来自R1外,其余3个QTL位点均由8908B提供增效基因。     

油菜和播娘蒿体细胞杂交创造油菜高油双低新种质

在甘蓝型双低油菜品种与播娘蒿原生质体融合杂种的自交后代群体中,实施定向选择和品质鉴定,在F3收获的247株中筛选获得9份含油量大于45%、芥酸含量小于0.5%、硫苷含量小于30μmol/g的高油双低甘蓝型油菜新种质,其中有2份是黄籽高油双低材料,这些材料的F4仍保持高含油量双低性状。在F4群体收获的905株中新选出了15份高含油量双低油菜新种质,其中有5份是黄籽高油双低材料。因此,通过播娘蒿与油菜原生质体融合及其后代选择可以获得高含油量优质油菜新种质。     

四个芸薹属物种硫苷含量与种类分析及特殊硫苷成分的种间导入

硫代葡萄糖苷(简称硫苷)是在十字花科植物中广泛存在的一类次级代谢物,在植物防御体系中发挥着重要的作用。本研究通过高效液相色谱(HPLC)技术,检测分析了十字花科芸薹属4个油用物种(甘蓝型油菜、埃塞俄比亚芥、芥菜型油菜、白菜型油菜)67个代表性品种3个器官(根、叶、种子)3个环境(2014年湖北武汉、2016年湖北武汉、2016年湖北襄阳)下的硫苷种类及含量。共检测到11种主要的硫苷,显著性检验结果显示:不同年份间硫苷含量分布差异极显著,但在同一年武汉与襄阳两个地点间差异不显著;不同物种间硫苷的类型差异明显;甘蓝型油菜、埃塞俄比亚芥不同品种间硫苷含量分布差异极显著,但受测的芥菜型油菜、白菜型油菜不同品种间硫苷含量分布差异不显著;同一个品种不同器官间的硫苷总量差异极显著;同一器官不同硫苷的含量差异也极为显著。这些结果表明硫苷在不同物种、不同器官、不同品种间都展现了丰富的变异,同时硫苷也受到了极显著的环境影响。每个物种或物种的不同器官内都有其代表性的特殊硫苷成分,如甘蓝型油菜中的2-羟基-3-丁烯基硫苷,埃塞比亚芥以及芥菜型油菜中的2-丙烯基硫苷,白菜型油菜中的3-丁烯基硫苷。其中一些特殊硫苷成分具有有益的生物学功能,它们的存在为通过种间杂交的方式将其向甘蓝型油菜中聚集提供了可能性。我们以在埃塞俄比亚芥与芥菜型油菜中所检测到的具有抗病功能的2-丙烯基硫苷为例进行展示,这种硫苷成分在高、中、低硫苷的甘蓝型油菜品种中含量都几乎为0,而在含有埃塞俄比亚芥与白菜型油菜亚基因组导入片段包含3个世代183个株系的再合成甘蓝型油菜(新型甘蓝型油菜)中,有170个株系检测到了显著高于品种材料的2-丙烯基硫苷成分,这说明了通过种间交流的方式向目标物种中导入特殊硫苷成分的可行性。以上这些结果将为分析利用芸薹属不同油用作物中的硫苷,并创建具有特殊硫苷成分的芸薹属作物提供信息和新的视角。     

不同季节黑杨萎蔫叶片挥发物的化学成分分析

采用水蒸气蒸馏法和气相色谱-质谱联用技术,分析黑杨不同季节的叶片,即嫩叶、成熟叶和衰老叶萎蔫后释放的气味物质成分.结果表明,嫩叶挥发物主要成分为顺-3-己烯醇、-甲基-1-戊醇和邻羟基苯甲醛,其相对含量分别为44.81%、21.85%和15.19%;成熟叶挥发物主要组分也是顺-3-己烯醇,相对含量28.71%,其次为邻羟基苯甲醛,相对含量10.35%.邻羟基苯甲醛为衰老叶挥发物的主要组分为,其相对含量为28.81%,其次是苯甲醇,相对含量15.06%;随着叶龄增长,挥发物中顺-3-己烯醇含量显著减少,而具有芳香性化合物种类及其含量显著增加.     

萝卜和白菜属间杂种硫甙基因表达与组分特征分析

硫甙是十字花科蔬菜中重要的次生代谢物质。在不同的十字花科作物中,硫甙的种类及其降解产物不同,从而产生了各自的特殊风味与生物功能。萝卜的主要硫甙4-甲基亚磺酰基-3-丁烯基硫甙(RAE)及其降解产物具有强力的抗癌活性;而白菜的主要硫甙2-羟基-3-丁烯基硫甙(PRO)的降解产物噁唑烷硫酮,被认为有致甲状腺肿大的副作用。本研究采用高效液相色谱分析法,分析了萝卜(‘36-2’)、白菜(‘Chiifu’)以及属间杂种(DH)中硫甙组分及其含量,共鉴定出13种硫甙。萝卜中的主要硫甙是4-甲硫基-3-丁烯基硫甙(RSA)和RAE;白菜中的主要硫甙为PRO、3-丁烯基硫甙(NAP)和吲哚-3-甲基硫甙(GBC);远缘杂交种的主要硫甙则有RAE、4-戊烯基硫甙(GBN)、NAP和PRO。通过与‘36-2’的比较转录组分析,发现DH中硫甙通路基因有14个下调表达和23个上调表达,其中萝卜特有基因RsIQD1.1和RsMYB34.2下调表达,而RsMYB34.1上调表达。值得注意的是萝卜中的RsFMOGS-OX2.1在DH中显著上调表达。通过与‘Chiifu’的比较分析,发现DH中硫甙通路基因有24个下调表达和15个上调表达,其中白菜特有基因BrIPMI-SSU3.1、BrIPMDH3.1和BrMAM3.1显著上调表达。这些基因在不同物种中的表达特点与其硫甙组分的变化有关。这在一定程度上证实远缘杂交可能通过改变后代的遗传组成和基因表达而改变其硫甙组成及含量,为以属间杂种作为桥梁种质实现物种间基因交流,创制高品质十字花科新种质奠定了理论基础和技术支撑。     

外源喷施萘乙酸对小白菜硫代葡萄糖苷的影响

本试验研究了不同浓度的萘乙（NAA）并结合单次喷施（NAA-1）和两次喷施（NAA．2）,对小白菜生长和硫代葡萄糖苷（简称硫苷）含量的影响。结果表明,与对照相比,不同的NAA处理浓度均显著增加了小白菜的鲜重。同时,NAA处理对总硫苷和单个硫苷含量产生了显著的影响。NAA-1处理时,总脂肪族硫苷、2．苯乙基硫苷和总硫苷在20mg·L-1时到达最大值;而总吲哚族硫苷在50mg·L-1时含量到达最高。NAA．2处理时,大部分单个硫苷和总硫苷在10mg·L-1处理即达到最大值。可见,在较低浓度两次喷施NAA试验中对大部分硫苷的诱导效果高于单次喷施NAA;但随着NAA处理浓度的提高,单次喷施NAA对硫苷的诱导效果较好。其中,吲哚-3-甲基硫苷含量在NAA-1处理,喷施50mg·L-1时显著高于其他处理：而2-苯乙基硫苷在单次和两次喷施NAA时,分别在20mg·L-1和10mg·L-达到了最大值。     

播娘蒿种子中几种硫苷的提取及鉴定

用石油醚对播娘蒿种子脱脂,热水提取,并采用DEAE sephadex A-25层析预处理,除去杂质,得到播娘蒿种子的硫苷提取液。利用高效液相色谱-电喷雾电离质谱(HPLC-ESI/MS)技术,并结合二级质谱分析,鉴定播娘蒿种子中主要硫苷的分子结构。鉴定出的6种硫苷成分的侧链结构分别为:3-丁烯基、3-甲硫丙基、苯甲基、5-氧代辛基、3-羟基-5-(甲基亚硫酰基)戊基、3-羟基-5-(甲基磺酰基)戊基。其中苯甲基硫苷具有抗癌作用。     

巴西橡胶树4-羟基-3-甲基-2-(E)-丁烯基-4-磷酸还原酶基因(Hb-HDR)的克隆及表达分析

4-羟基-3-甲基-2-(E)-丁烯基-4-磷酸还原酶(4-hydroxy-3-methyl-2-(E)-butenyl-4-diphosphatereductase,HDR)是异戊烯基焦磷酸合成途径之一甲基赤藓糖磷酸(methylerythritol phosphate,MEP)途径中的最后一个酶,催化4-羟基-3-甲基-(2E)-丁烯基-4-磷酸生成异戊烯基焦磷酸.根据植物HDR的同源序列设计引物,通过RT-PCR结合RACE的方法在橡胶树中获得了与其相应的HDR基因,命名为HbHDR.序列分析表明HbHDR长1 627 bp,编码462个氨基酸,属于LYTB家族,该氨基酸序列与烟草、长春花、胡黄连、拟南芥、银杏和火炬松的HDR同源性为79.1%、78.4%、76.5%、75.3%、72.2%和70.9%.半定量RT-PCR结果显示,乙烯诱导胶乳HbHDR的表达.     

环境对植物芥子油苷代谢的影响

芥子油苷是一类含氮、含硫的植物次生代谢物质,主要分布于十字花科植物.芥子油苷及其降解产物具有多种生化活性,近年来人们更多地关注芥子油苷代谢与植物生存环境的相互作用以及与其它物质代谢途径的相互联系.介绍了温度、光、水分、硫营养、CO2浓度以及重金属污染等非生物环境对芥子油苷代谢影响的研究概况.     

Suppressive Effect of Yamato-mana (Brassica rapa L. Oleifera Group) Constituent 3-Butenyl Glucosinolate (Gluconapin) on Postprandial Hypertriglyceridemia in Mice

We examined the bioactivity of Yamato-mana (Brassica rapa L. Oleifera Group) constituent glucosinolates and found that 3-butenyl glucosinolate (gluconapin) decreased the plasma triglyceride gain induced by corn oil administration to mice. However, phenethyl glucosinolate (gluconasturtiin) had little effect. 2-Propenyl glucosinolate (sinigrin) also reduced the plasma triglyceride level, which suggests that alkenyl glucosinolates might be promising agents to prevent postprandial hypertriglyceridemia.     

Studies on glucosinolate degradation in Lepidium sativum seed extracts

Analysis of Lepidium sativum seeds showed the presence of allyl, 2-phenethyl and benzyl glucosinolates, the first two being reported for the first time from this source. The effects of temperature, pH of the extraction medium and the length of time allowed for autolysis were assessed on the benzyl glucosinolate degradation products in seed extracts. In particulàr benzyl thiocyanate was not produced at higher temperatures but at ambient and lower temperatures it exceeded isothiocyanate. Nitrile was always the major product under the conditions studied, ever at pH levels as high as 7.4. Five new possible benzyl glucosinolate degradation products were detected and evidence is presented that benzaldehyde and benzyl alcohol could be secondary products formed thermally from isothocyanate and thiocyanate, respectively. Benzyl mercaptan and benzyl methyl sulphide also appear to be thermally produced.     

Effects of metal ions on benzylglucosinolate degradation in Lepidium sativum seed autolysates

The effects of varying concentrations of Fe²⁺ (5 × 10^?5 ?5 × 10^?1 M) on benzylglucosinolate degradation in Lepidium sativum seed autolysates were investigated. Increased glucosinolate decomposition was observed over the whole range with a maximum effect at ca 6 × 10^?3 M Fe²⁺, at which point glucosinolate degradation was more than three times that obtained in the absence of added Fe²⁺ . Nitrile formation was especially enhanced in the presence of all concentrations of Fe²⁺ studied, and maximum amounts were obtained at ca 6 × 10^?3 M Fe²⁺ when a more than four-fold increase over quantities produced in the absence of Fe²⁺ was observed. Thiocyanate formation was also promoted with a maximum at ca 4 × 10^?3 M Fe²⁺, but isothiocyanate production was considerably reduced in allcases. It is suggested that Fe²⁺ inhibits isothiocyanate formation by interfering with the availability of ascorbic acid which is a proven co-factor for most thioglucosidase isoenzymes, but that an Fe²⁺-ascorbate complex might then be responsible for promoting enzymic production of nitrile. The effects of a limited range of concentrations of Fe³⁺ and Cu⁺ were also studied, and results related to those for Fe²⁺. The relevance of the findings to natural systems and to glucosinolate-containing foods is briefly discussed.     

3-Hydroxypropylglucosinolate,a new glucosinolate in seeds of Erysimum hieracifolium and Malcolmia maritima

Glucosinolates from seed meals of Erysimum hieracifolium and Malcolmia maritima were treated with thioglucosidase (EC 3.2.3.1), and the resultant aglucon products investigated. A major product from E. hieracifolium was 3-hydroxypropyl isothiocyanate from the novel precursor 3-hydroxypropylglucosinolate. Other aglucon products were 3-methylsulfonylpropyl, 3-methylsulfinylpropyl, 3-methylthiopropyl, and allyl isothiocyanates. The aglucon products from M. maritima seed meal included 3-hydroxypropyl isothiocyanate, in addition to the previously known 3-methylsulfonylpropyl and 3-benzoyloxypropyl isothiocyanantes.     

Sulphur Deficiency Causes a Reduction in Antimicrobial Potential and Leads to Increased Disease Susceptibility of Oilseed Rape

The reduction of atmospheric sulphur dioxide pollution is causing increasing problems of sulphur deficiency in sulphur‐demanding crop plants in northern Europe. Elemental sulphur and many sulphur containing compounds such as cysteine‐rich antifungal proteins, glucosinolates (GSL) and phytoalexins play important roles in plant disease resistance. The aim of this work was to analyse the effect of inadequate sulphur supply on disease resistance of oilseed rape (Brassica napus). Compared with fertilized oilseed rape, healthy looking S‐deficient plants showed increased susceptibility to the blackleg fungus Leptosphaeria maculans, to the generalist necrotroph Botrytis cinerea and to the oomycete Phytophthora brassicae. To analyse possible causes of the increased disease susceptibility of S‐deficient plants, protein extracts and methanolic extracts of secondary metabolites of plants grown with and without adequate sulphur supply were tested for antimicrobial activity. None of the protein extracts showed antimicrobial activity. However, extracts containing secondary metabolites from normally grown plants showed a strong antimicrobial activity in in vitro tests with various fungal and bacterial pathogens. This activity was almost totally lost in extracts derived from S‐deficient plants. The antimicrobial activity did not appear to be based on the activity of phytoalexins because it was present in healthy plants and was not increased by a previous inoculation with Botrytis cinerea. The loss of antifungal activity in S‐deficient plants correlated with a strong reduction of various GSL, thus suggesting a reduced level of GSL as a possible cause of the reduced antimicrobial potential. However, limited tests of commercially available GSL or their degradation products did not demonstrate a causal link. Our results show that S‐deficiency of oilseed rape negatively affects disease resistance and suggest that this effect is at least partially caused by a reduction of sulphur‐dependent phytoanticipins.     

茉莉酸类化合物在植物防卫反应中的作用

茉莉酸类化合物(jasmonate,JA)是诱导植物防卫反应的重要信号分子。JA不仅是系统素信号转导途径的重要组分,而且在植物长距离伤信号转导中发挥重要作用。JA还能单独或与其他激素相互作用调节与植物防卫反应相关的次生代谢物质芥子油苷的生物合成,从而影响植物的防卫反应。现对JA在植物防卫反应中的作用进行综述,并对今后这一领域的研究进行了展望。     

机械损伤对拟南芥莲座叶芥子油苷含量和组成的影响

植物可以利用体内次生代谢产物的变化来抵御昆虫取食和机械损伤.芥子油苷是拟南芥的主要次生代谢产物.通过剪刀剪取叶片(40%面积)对温室培养的拟南芥幼苗莲座叶进行机械损伤处理,观察机械损伤后8个时间点拟南芥叶片中不同种类芥子油苷含量和组合模式的变化.结果表明机械损伤后3 h叶片中芥子油苷总含量开始明显上升,脂肪族和吲哚族芥子油苷含量在损伤后3 h也都显著高于损伤前.在检测到的12种芥子油苷中,4-甲基亚磺酰丁基芥子油苷(4-methylsulphinylbutyl GS,4MSOB)的含量最多,占芥子油苷总量的48.5%,并且在损伤3 h后含量增加.4MSOB含量的变化成为影响莲座叶中芥子油苷组合模式的主导因素.其它各种芥子油苷在损伤后不同时间点的变化也存在差异.