菘蓝特性及栽培措施的研究

板蓝根和大青叶是临床上广泛应用的两味中药,但由于市场上品种混杂,真假难辨,直接影响了疗效。为发展正品药材,作者对原植物菘蓝的特性及栽培措施进行了研究。     

3种十字花科植物的核型研究

３种十字花科植物的核型研究肖成汉,赵建伟,何凤仙（华中农业大学武汉４３００７０）关键词十字花科,菘蓝,芝麻菜,墙生二行芥,核型ＳＴＵＤＩＥＳＯＮＫＡＲＹＯＴＹＰＥＳＯＦＴＨＲＥＥＳＰＥＣＩＥＳＩＮＣＲＵＣＩＦＥＲＡＥ￥ＸｉａｏＣｈｅｎｇｈａｎ;Ｚｈａ...     

缺氮和复氮对菘蓝幼苗生长及氮代谢的影响

对基质育苗后水培的菘蓝进行缺氮与复氮处理,分析其生长情况及氮代谢产物含量的变化,探讨缺氮和复氮对菘蓝幼苗生长及氮代谢的影响,以提高菘蓝产量和品质以及栽培过程中的氮素利用效率。结果显示:(1)正常供氮条件下,菘蓝幼苗的叶绿素含量、谷氨酰胺合成酶(GS)活性、硝态氮含量、靛玉红含量为最高,而其株高、主根直径、根的鲜重与干重、叶的鲜重与干重、根系活力均最小。(2)缺氮处理增加了菘蓝幼苗的主根直径和根干重,提高其根系活力和硝酸还原酶(NR)活性,促进游离氨基酸在叶中的积累;但降低了GS的活性,也降低了叶中硝态氮、可溶性蛋白、靛玉红及根中游离氨基酸的含量;缺氮对叶中靛蓝的含量无明显影响。(3)复氮处理增加了菘蓝幼苗的株高、主根长、根鲜重、叶鲜重、叶干重,提高了其根系活力,降低了NR和GS的活性;与对照相比,复氮降低了叶中硝态氮含量,提高了叶中可溶性蛋白、靛蓝及根中游离氨基酸的含量,但对叶中游离氨基酸和靛玉红含量影响较小。研究表明,缺氮后再复氮有利于菘蓝幼苗叶的生长,同时有利于增加其叶内靛蓝含量,从而提高其产量和品质。     

氮素形态及配比对5个种源菘蓝的生物碱类成分和多糖的影响

以5份不同种源的菘蓝为材料,采用田间小区试验,设置不施氮(CK)、硝态氮(NO3--N)、铵态氮(NH4+-N)、NH4+-N/NO3--N=75/25、NH4+-N/NO3--N=50/50、NH4+-N/NO3--N=25/75和酰胺态氮等7个处理,分析比较了不同种源植株的靛蓝、靛玉红和总生物碱含量、(R,S)-告依春及多糖含量等指标的差异,为菘蓝栽培生产中氮素的高效利用提供理论参考。结果表明:氮素处理有利于提高山西运城菘蓝和陕西商洛菘蓝叶内靛蓝含量,以及安徽亳州菘蓝和陕西商洛菘蓝叶内的总生物碱含量;NH4+-N/NO3--N=50/50处理对山西运城菘蓝,以及酰胺态氮处理对山西运城菘蓝和陕西商洛菘蓝叶内生物碱类成分的积累均有促进作用;与对照相比,氮素处理亦能有效地提高甘肃张掖菘蓝和陕西商洛菘蓝根内的(R,S)-告依春及安徽亳州菘蓝根内的多糖含量;安徽阜阳菘蓝(R,S)-告依春含量在任一氮处理下均远远高于其他种质菘蓝。研究表明,不同种源菘蓝对氮素处理的响应存在较大的差异,建议生产中综合考虑菘蓝的来源和需肥规律,采用经济有效的施氮组合,以提高其活性成分含量。     

菘蓝不同器官脂溶性成分的GC-MS分析

采用索氏提取法对菘蓝根、茎、叶、花和幼果等不同器官的脂溶性成分进行提取,利用气相色谱-质谱联用(GC-MS)技术对其脂溶性成分进行鉴定,并运用面积归一化法确定相对含量,以明确菘蓝的化学物质基础,为综合开发利用菘蓝资源提供依据。结果显示:从菘蓝的根、茎、叶、花和幼果中分别鉴定出24、23、21、18和23种脂溶性成分;其中,十二烷、十三烷、十四烷、2,4-二叔丁基苯酚和棕榈酸为各器官的共有成分;不同器官中脂肪酸含量依次为:花(80.91%)>根(60.56%)>幼果(21.93%)>茎(17.45%)>叶(11.28%)。除花中的饱和脂肪酸含量高于不饱和脂肪酸外,其他器官中不饱和脂肪酸的含量均较高,表明菘蓝不同器官中脂溶性成分的组成及含量存在一定差异。     

菘蓝IiCYP79F1基因的克隆与表达分析

该研究以菘蓝叶片为材料,采用RT PCR方法克隆菘蓝IiCYP79F1基因,并对其进行生物信息学与表达模式分析。结果表明：（1）成功获得IiCYP79F1基因的gDNA全长（2 109 bp）,包含3个外显子及2个内含子,ORF全长为1 626 bp,编码541个氨基酸（GenBank登录号为KY774689.1）;生物信息学分析显示,IiCYP79F1蛋白的二级结构主要由α 螺旋（43.81%）、无规则卷曲（35.49%）、延伸链（13.68%）和β 转角（7.02%）组成;其氨基酸序列与西兰花、欧洲油菜、芜菁和芝麻菜的相似性较高,其蛋白与芝麻菜的亲缘关系最近。（2）qRT PCR分析显示,IiCYP79F1基因呈时空特异性表达,且在茎中与幼苗期高表达;MeJA、Ag⁺、葡萄糖和机械损伤处理均能有效促进IiCYP79F1基因的表达,而SA和低温处理则具有明显的抑制作用。该研究结果为进一步探讨IiCYP79F1在菘蓝芥子油苷生物合成中的作用奠定了基础,也为培育高芥子油苷含量的菘蓝新种质提供了新思路。     

Interspecific water use strategies of a Juglans regia and Isatis tinctoria/Senna tora agroforestry

Aims Understanding the interspecific water relations is important for designing agroforestry systems. The objective of this study was to determine the water use strategies of component species in a walnut (Juglans regia)-woad (Isatis tinctoria)/sicklepod (Senna tora) agroforestry system.Methods Water sources of component species in a walnut-woad/sicklepod agroforestry system were investigated with the technique of stable deuterium isotope tracing at a site of hilly area in Northern China during 2012-2013.Important findings Results showed that the soil water content in the agroforestry system was 26.74% and 7.93% greater than in the pure woad field in the first half year, and 17.39% and 13.65% greater than in the pure sicklepod field in the second half year (sicklepod growth period), in 2012 and 2013, respectively. The lowest water content was found in the middle of tree rows, and the highest water content was found in the northern side of tree rows or under the trees. In the soil layers measured, the pure woad and pure sicklepod systems had greater hydrogen stable isotope ratios (δ D value) of soil water than in the agroforestry system. During the period of woad growth, more than half of the water absorbed by walnut was from the deeper soil layer (30-80 cm). In contrast, the walnut trees mainly utilized shallow layer (0-30 cm) soil water during the period of sicklepod growth. These findings suggest that walnut has a two-state root system: during the period of woad growth, shallow roots of walnut are not active when soil is dry whereas the sicklepod growth occur in rainy season, and the shallow roots of walnut are active and utilize more shallow soil water supplemented by rainwater. More than 85% of water used by both the woad and the sicklepod were from the shallow layer soil. At the seedling stage, the roots of woad, cannot grow into the deeper soil layer, and the absorbed water is completely from the shallow layer in the pure woad system. However, 5.7% of the water absorbed by the intercropped woad was from the deeper soil layer in 2012, and the proportion increased further (9.7%) in the following year when there was less precipitation. The results confirmed that hydraulic lift effect of walnut occurred on shallow layer crop in dry season, and this effect become greater under drier conditions. Therefore, deeper roots of walnut improved water condition in the walnut- woad/sicklepod agroforestry systems compared to pure crop systems. The walnut mainly utilized water from the deeper layer to avoid water competition with the shallow layer. In the dry season, crops benefited from the water provided by walnut roots through hydraulic lift. Walnut and intercropped plants exhibited water facilitation in the agroforestry systems, suggesting that this configuration is a suitable practice in this area.     

菘蓝二倍体及其同源四倍体遗传差异的ISSR分析

以1个菘蓝二倍体及其10个同源四倍体株系为材料,利用19个随机扩增ISSR引物分析其遗传差异性,为菘蓝多倍体诱变的基因表达调控以及遗传改良提供依据。结果显示:菘蓝二倍体与其同源四倍体及四倍体之间的ISSR多态性有明显差异,除主要遗传位点相同外,有些四倍体株系扩增条带数多于二倍体,有些四倍体株系扩增条带数少于二倍体,四倍体株系间亦有多态位点;11个株系共扩增111条多态性条带,多态性达55.22%。聚类分析显示,不同四倍体株系与二倍体的遗传差异大小亦不同。研究表明,菘蓝二倍体与其同源四倍体具有中等偏高的遗传差异性。     

华北低丘山区果药复合系统种间水分利用策略

了解林农复合系统的种间水分关系至关重要.该文通过稳定氘同位素研究了华北低丘山区核桃(Juglans regia)-菘蓝(Isatis tinctoria)/决明(Senna tora)复合系统各组分的水分来源, 试图明确该果药复合系统的种间水分利用策略, 为该区林农配置模式的选择提供理论依据.研究结果表明: 果药复合系统的土壤含水量明显高于单作菘蓝和单作决明地块, 在2012年,2013年上半年比单作菘蓝高26.74%和7.93%, 在下半年比单作决明高17.39%和13.65%.在果药复合系统内部, 土壤含水量以核桃树行中间位置的最低,树行北侧和树下最高.在各个土层深度, 单作系统的土壤水氢稳定同位素比率(δD值)均比复合系统的高.在菘蓝生长时期的春旱期, 复合系统中核桃的大部分水分来源于30-80 cm深层土壤水, 表明此时期核桃表层根系活性不高; 而决明生长时期正值雨季, 此时核桃优先利用雨水补充的0-30 cm浅层土壤水,表层根系活性增强.在任何生长时期, 菘蓝和决明85%以上的水分都来自浅层土壤水.在菘蓝苗期, 其根系尚未扎入深层土壤中, 单作菘蓝的水分完全来源于浅层土壤, 而在2012年间作菘蓝却有5.7%的水分来自于深层土壤, 在更为干旱的2013年该比例上升到9.7%, 该结果证实了核桃在旱季存在"水力提升"作用, 供浅根系作物吸收利用, 并且越干旱, 该水力提升作用越强.在华北低丘山区核桃-菘蓝/决明复合系统中, 深根性核桃改善了复合系统的土壤水分状况, 在旱季主要利用深层土壤水以避开与浅层作物的水分竞争,并能将深层土壤水提升至浅层土壤供菘蓝吸收利用, 核桃与两种药材表现为水分互利关系, 因而该模式适合在该地区发展.     

菘蓝花药培养及单倍体的诱导

探讨菘蓝花药处于单核晚期的形态指标,并以适宜发育时期的花药为外植体,进行花药培养及单倍体诱导。实验结果表明,4℃低温处理2d后,在含有6-BA0．5mg·L-1和NAA1．0mg·L-1。的Ms培养基上,花药愈伤组织的诱导率为23．35％;将其转接到Ms附加6．BA1．0mg·L-1,NAA0．5mg·L-1的分化培养基上,80．00％以上的愈伤组织可以诱导产生不定芽;再将分化出的试管苗转接到1／2MS＋NAA1．0mg·L-1的生根培养基上,3d左右即可获得完整植株。经叶边缘压片检查染色体数目,花药培养所得的弱小绿苗为单倍体植株。