一种促进麻疯树再生不定芽芽条生根的方法 An Jatropha regeneration of adventitious buds buds rooting method of promoting

技术领域 FIELD

[0001] 本发明涉及植物生物技术领域，具体地，涉及一种促进麻疯树再生不定芽芽条生根的方法。 [0001] The present invention relates to the field of plant biotechnology, in particular, to a jatropha regeneration of adventitious buds buds promote rooting method.

背景技术 BACKGROUND

[0002] 伴随着全球对化石燃料的需求日益增长，存储的化石燃料即将耗尽，人们越来越关注可再生的生物柴油。 [0002] With the growing global demand for fossil fuels, fossil fuel is about to run out of storage, there is a growing focus on renewable biodiesel. 在可产生生物柴油的候选植物中，属于大戟科的麻疯树(JatrophacurcasL.)具有明显的优势,它的种子含油量高,种仁含油量可达40%~60%,同时，麻疯树油中含有活性成分多，如毒蛋白、麻疯酮等有着重要的农药和医药价值。 In the candidate can produce biodiesel plant belonging Euphorbiaceae Jatropha (JatrophacurcasL.) Has obvious advantages, its high seed oil content, seed kernel oil content of up to 40% to 60%, while, leprosy tree oil contains more active ingredients, such as toxic protein, ketones such as leprosy has important value for pesticides and pharmaceuticals.

[0003] 然而，大力推广麻疯树的种植却面临一系列问题，虽然麻疯树种子中含油量高，但种子产量不高；种油中活性成分多，但仍需要改变油的成分才能直接替代化石燃料；麻疯树对环境要求较高，耐寒能力弱，分布区域较窄。 [0003] However, to promote the cultivation of Jatropha has faced a series of problems, although Jatropha seeds high oil content, but the seed yield is not high; the active ingredient in more kinds of oil, but still need to change the composition of the oil to direct alternative to fossil fuels; Jatropha higher environmental requirements, cold weak, narrow distribution area. 麻疯树属包括175个种，可以通过种间杂交引入优良性状，但所需周期长，不能够获得特异的外源基因，故主要通过基因转化改变遗传背景。 Jatropha, including 175 kinds, can be introduced through interspecific hybridization good traits, but the long period required, can not obtain specific exogenous gene, it is mainly by changing the genetic background genetic transformation. 高频率的植株再生体系是基因转化的基础。 Regeneration System is the foundation of the high frequency of genetic transformation.

[0004] 利用组织培养技术对植物进行再生或再生克隆，通常包含两个关键步骤。 [0004] The use of tissue culture techniques for cloning plants regenerated or recycled, usually contains two key steps. 第一步是从外植体诱导产生不定芽。 The first step is to induce adventitious buds from explants. 第二步是促使不定芽芽条生根，以备移栽。 The second step is to promote rooting of adventitious buds buds to prepare for transplanting. 对于麻疯树，现有技术中已有很多报道研究其外植体诱导产生不定芽；而至于如何促使这些不定芽芽条良好生根，则很少有人研究。 For Jatropha, the prior art has been widely reported study outer explants induced adventitious buds; but as to how to promote these good rooting of adventitious buds buds, it has seldom been studied.

[0005] 现有技术中，迄今为止已建立起来的诱导麻疯树再生不定芽芽条生根的方法一般都是在生根培养基中不添加激素或者只添加一定浓度的生长素，现有方法存在生根效率低、再生不定根的质量较差、所获得再生植株生长状态不佳、整个培养周期较长等缺点，难以满足实际应用的需要。 [0005] The prior art has so far established Jatropha induce regeneration of adventitious buds buds rooting methods are generally not in a rooting medium added hormones or just add a certain concentration of auxin, existing methods exist Rooting low efficiency and poor quality of regeneration of adventitious roots, and the poor get regenerated growth state, the longer the entire culture period and other shortcomings, it is difficult to meet the needs of practical application.

[0006] L-谷氨酰胺是一种条件必需性氨基酸，在生命活动中起着重要作用。 [0006] L- Glutamine is a conditionally essential amino acid, plays an important role in life activities. 它是构成蛋白质的氨基酸，又是合成含氮生物物质的氮源，与组织生长和修补有着密切的关系。 It is a protein amino acids, but also the synthesis of nitrogen-containing biomass nitrogen, and tissue growth and repair are closely related. 在不同组织中谷氨酰胺具有不同的代谢作用，起着重要的生理作用。 Glutamine has a different metabolic effects in different tissues, plays an important physiological role. 目前仅有一篇论文报道称，L-谷氨酰胺对石斛兰的生根有负面影响，在培养基中添加16mg/L的谷氨酰胺对石斛兰的生根和生长起抑制作用。 Currently, only a paper reported, L- glutamine have a negative effect on rooting Dendrobium, adding 16mg / L of glutamine on Dendrobium rooting and growth in the medium inhibited.

发明内容 SUMMARY

[0007] 本发明为了克服现有技术麻疯树不定芽芽条生根率低、生根质量差、培养周期长的缺陷，提供一种促进麻疯树不定芽芽条生根的方法。 [0007] The present invention to overcome the prior art Jatropha adventitious buds buds rooting rate, rooting poor quality, develop long cycle defects, provide a promotion of Jatropha adventitious buds buds rooting method. 采用本方法可以在较短时间内就使得再生不定芽芽条的生根率达到较高值，因此可显著缩短再生培养周期；同时减少了生根过程中常见的再生植株叶片发黄现象，提高了所获得再生植株的质量，使麻疯树相关生物技术育种的工作效率得到相应的大幅度的提高。 This method can be adopted in a relatively short period of time so that the regeneration of adventitious rooting rate Buds bar reaches a higher value, it can significantly shorten the regeneration cycle training; while reducing the rooting process commonly regenerated plants leaf yellowing and improve the access to quality regenerated plants, so the efficiency of Jatropha related biotechnology breeding has been a corresponding increase greatly.

[0008] 本发明通过以下技术方案予以实现上述目的: [0008] The present invention is to be realized by the following technical scheme above object:

[0009] 一种促进麻疯树再生不定芽芽条生根的方法，将不定芽芽条接种在含有4~32mg/LL-谷氨酰胺的生根培养基上培养10~40天。 [0009] An article promoting adventitious rooting method Buds Jatropha regeneration, the adventitious buds bud inoculation training 10 to 40 days in the rooting medium containing 4 ~ 32mg / LL- glutamine.

[0010] 所述L-谷氨酰胺母液的配制方法为:准确称取一定量的L-谷氨酰胺粉末，用去离子水充分溶解后，再用去离子水定容，配制成2mg/mL的L-谷氨酰胺母液。 [0010] The L- glutamine liquor preparation method: Accurately weigh a certain amount of L- glutamine powder, after fully dissolved in deionized water to volume with deionized water to prepare a 2mg / mL The L- glutamine mother liquor. 使用前用0.22微米的水系滤膜对已配制好的L-谷氨酰胺母液进行过滤灭菌。 Before using a 0.22 micron filter water system have been formulated L- glutamine liquor filter sterilization. 使用时，再通过加入无菌水将L-谷氨酰胺母液配置成4、8、16、32、64、128mg/L的L-谷氨酰胺工作液。 When used, by the addition of sterile water and then L- glutamine mother liquor configured 4,8,16,32,64,128mg / L of L- glutamine working fluid.

[0011] 所述不定芽芽条的获取方法为:从培养瓶中取出具有长度大于Icm且至少有2个伸展叶片的不定芽的外植体，用灭过菌的手术刀对伸长的再生不定芽进行横向切割(切割的方向与伸长的再生不定芽的中轴相垂直，切口要平齐)，切去其他部分，获取长度约为Icm的不定芽芽条。 [0011] The adventitious buds buds acquisition method is as follows: Remove the explant has a length greater than Icm and at least two extension leaves adventitious buds from culture flasks with a sterile scalpel elongation regeneration adventitious buds horizontal cut (cutting direction and regeneration of adventitious buds elongated axis perpendicular incision should flush), cut the other section for a length of about Icm adventitious buds buds.

[0012] 优选地，将不定芽芽条接种在含有8~32mg/LL_谷氨酰胺的生根培养基上培养10~40天。 [0012] Preferably, the adventitious shoot buds were inoculated 10 to 40 days in culture on a rooting medium containing 8 ~ 32mg / LL_ glutamine.

[0013] 更优选地，将不定芽芽条接种在含有16mg/LL_谷氨酰胺的生根培养基上培养10天或将不定芽芽条接种在含有8~32mg/LL-谷氨酰胺的生根培养基上培养20~40天。 [0013] More preferably, the adventitious shoot buds seed culture for 10 days on a rooting medium containing 16mg / LL_ glutamine or adventitious buds buds inoculated containing 8 ~ 32mg / LL- glutamine rooting the culture medium 20 to 40 days.

[0014] 最优选地，将不定芽芽条接种在含有16mg/LL_谷氨酰胺的生根培养基上培养10~40天。 [0014] Most preferably, the adventitious shoot buds were inoculated 10 to 40 days in culture on a rooting medium containing 16mg / LL_ glutamine.

[0015] 优选地，所述接种的方式为竖插方式，即使芽条的中轴与培养基表面相垂直，芽条的形态学下端插入培养基中的深度为0.2~0.4cm。 [0015] Preferably, the inoculated way for vertical interpolation mode, even if the central axis of the medium perpendicular to the surface of shoots, shoot morphology of the lower end of the medium insertion depth of 0.2 ~ 0.4cm.

[0016] 优选地，所述培养的条件为光照强度为2000~25001x，光照时间为12~16小时/天，培养温度为25±1°C。 [0016] Preferably, the conditions for the cultivation of the light intensity of 2000 ~ 25001x, illumination time is 12 to 16 hours / day, the culture temperature was 25 ± 1 ° C.

[0017] 优选地，所述生根培养`基还含有0.3mg/L吲哚丁酸，30g/L蔗糖，100mg/L肌醇，5g/L琼脂和大量元素减半的MS培养基成分。 [0017] Preferably, the rooting `yl further contains 0.3mg / L indole butyric acid, 30g / L sucrose, 100mg / L myo-inositol, 5g / L agar and macronutrient halving MS medium components.

[0018] 本发明的有益效果: [0018] The beneficial effects of the present invention:

[0019] 本发明主要是改变了传统诱导麻疯树再生不定芽芽条生根的方法，即在诱导不定根再生的培养基中添加一定浓度的L-谷氨酰胺，由此促使部分细胞以更高的效率直接再分化形成更多的不定根。 [0019] The present invention is to change the traditional method primarily induce regeneration of adventitious buds Jatropha rooting buds, that add a certain concentration of L- glutamine in the medium to induce regeneration of adventitious roots, thereby causing some cells to higher The efficiency of the direct re-differentiate into more adventitious roots. 此外，采用本方法可以在较短时间内就使得再生不定芽芽条的生根率达到较高值，因此可显著缩短再生培养周期；同时减少了生根过程中常见的再生植株叶片发黄现象，提高了所获得再生植株的质量，使麻疯树相关生物技术育种的工作效率得到相应的大幅度的提闻。 In addition, the use of this method in a relatively short period of time so that the regeneration of adventitious rooting rate Buds bar reaches a higher value, it can significantly shorten the regeneration cycle training; while reducing the rooting process commonly regenerated plants yellow leaves, and enhanced the quality of regenerated plants obtained, so that the efficiency of Jatropha related biotechnology breeding corresponding substantial mention the smell.

[0020] 说明书附图 [0020] The accompanying drawings

[0021] 图1.麻疯树再生不定芽芽条在6种常见的传统生根培养基中进行不定根诱导培养30 天后的效果；A: MS; B: 1/2MS; C: l/2MS+lmg/LIBA+lmg/LIAA+2mg/LNAA; D: l/2MS+lmg/LNAA;E:l/2MS+lmg/LIAA;F:l/2MS+lmg/LIBA (bar=Icm)D [0021] Figure 1. Jatropha regeneration of adventitious buds induced adventitious root buds were cultured for 30 days results in six kinds of common traditional rooting medium; A: MS; B: 1 / 2MS; C: l / 2MS + lmg / LIBA + lmg / LIAA + 2mg / LNAA; D: l / 2MS + lmg / LNAA; E: l / 2MS + lmg / LIAA; F: l / 2MS + lmg / LIBA (bar = Icm) D

[0022] 图2.不同浓度L-谷氨酰胺对麻疯树再生不定芽芽条生根的作用效果与再生植株生长的效果；A: Omg/L; B: 4mg/L; C: 8mg/L; D: 16mg/L; E: 32mg/L; F: 64mg/L; G: 128mg/LL_ 谷氨酰胺的生根培养基中培养30d后的生根效果图；H:再生植株在土壤中继续生长的效果图(bar=lcm)。 [0022] Figure 2. Different concentrations of L- glutamine on Jatropha regeneration of adventitious buds buds rooting effect plant growth and regeneration effects; A: Omg / L; B: 4mg / L; C: 8mg / L ; D: 16mg / L; E: 32mg / L; F: 64mg / L; G: 128mg / LL_ glutamine rooting medium rooting renderings after 30d; H: regenerated plants continue to grow in the soil renderings (bar = lcm).

[0023] 图3.麻疯树再生不定芽芽条在添加16mg/LL_谷氨酰胺的生根培养基中不同培养时间下的生根效果与再生植株生长的效果;八和£:10(1;8:20(1;(::30(1;0，F和G:40d后的生根效果图；H:再生植株在土壤中继续生长的效果图(bar=lcm)。[0024] 图4.麻疯树再生不定芽芽条在添加了L-谷氨酰胺的生根培养基中培养10天后的效果；A:8mg/LL-谷氨酰胺；B: 16mg/LL-谷氨酰胺；C:32mg/LL_谷氨酰胺(bar=lcm)。 [0023] Figure 3. Jatropha regeneration of adventitious buds buds adding 16mg / LL_ glutamine rooting medium rooting effect of different culture and regeneration of plants grown under time; eight and £: 10 (1; 8:20 (1; (:: 30 (1; 0, F and G: after rooting Figure 40d; H:. regenerated plants grown in the soil continue renderings (bar = lcm) [0024] FIG. Jatropha regeneration of adventitious buds buds added L- glutamine in rooting medium effect for 10 days; A: 8mg / LL- glutamine; B: 16mg / LL- glutamine; C: 32mg / LL_ glutamine (bar = lcm).

具体实施方式 DETAILED DESCRIPTION

[0025] 下面结合附图和具体实施例进一步详细说明本发明。 [0025] below with reference to the accompanying drawings and the specific examples further illustrate the present invention. 除非特别说明，实施例中采用的试剂和方法为本领域常规使用的试剂和方法。 Unless otherwise specified, reagents and methods and reagents employed in the method of Example conventionally used in the art.

[0026] 实施例1 [0026] Example 1

[0027] 采用传统方法诱导麻疯树再生不定芽芽条生根的效果 [0027] Jatropha using traditional methods to induce regeneration of adventitious buds buds rooting effect

[0028] S1.再生不定芽芽条的获取:从培养瓶中取出具有长度大于Icm且至少有2个伸展叶片的不定芽的外植体，用灭过菌的手术刀对伸长的再生不定芽进行横向切割(切割的方向与伸长的再生不定芽的中轴相垂直，切口要平齐)，切去其他部分，获取长度约为Icm的不定芽芽条。 . [0028] S1 adventitious buds bud regeneration of the acquisition: removed from culture flasks with a length greater than Icm adventitious buds and at least two extension leaves explants with a sterile scalpel to the elongation of the regeneration of adventitious lateral buds cutting (cutting direction and elongation of adventitious buds regeneration perpendicular axis, the incision should be flush), cut the other section for a length of about Icm adventitious buds buds.

[0029] S2.将SI获取的芽条以竖插方式(使芽条的中轴与培养基表面相垂直，芽条的形态学下端插入培养基中的深度为0.2~0.4cm。)接种至6种传统生根培养基上培养30天，所获得实验结果如表1和图1所示。 [0029] S2. The SI acquired bud bar to vertical interpolation method (so buds of the axis perpendicular to the surface of the medium, bud morphology of the lower end of the insertion depth of the medium is 0.2 ~ 0.4cm.) Inoculated 6 kinds of traditional cultured on rooting medium for 30 days, the obtained results are shown in Table 1 and Figure 1.

[0030] 表1采用传统方法诱导麻疯树再生不定芽芽条生根的效果 [0030] Table 1 using the traditional method of inducing regeneration of adventitious buds Jatropha buds rooting effect

[0031] [0031]

[0032] 注:数据采用SPSSStatisticsl7.0统计分析软件进行方差分析和邓肯多重比较(P兰0.05)，数据后字母不同表示处理间差异显著。 [0032] Note: The data using statistical analysis software SPSSStatisticsl7.0 ANOVA and Duncan's multiple comparison (P lan 0.05), after the data is different letters indicate significant differences between treatments. 生根率(％)=(再生出不定根的外植体数/总外植体数)X 100% ;平均每个外植体的根数(条)=再生不定根总数/再生出不定根的外植体数；平均根长(cm)=再生不定根长度的平均值。 Rooting rate (%) = (regeneration of adventitious root explants / total number of explants) X 100%; the average root number (bar) = each explant regeneration of adventitious roots total / regeneration of adventitious root explants count; average root length (cm) = average of adventitious root regeneration length.

[0033] 从表1可知，在6种诱导芽条生根培养基中，生根效果最好的是添加了lmg/LIBA的1/2MS培养基，芽条在这种培养基上培养30天后，其生根率为16.10%，平均每个芽条上的根数为2.58条，平均根长为2.51cm ;芽条在无激素培养基(MS和1/2MS)和添加三种不同生长素的培养基(l/2MS+lmg/LIBA+lmg/LIAA+2mg/LNAA)中都不能产生不定根，说明生长素可能是诱导芽条生根所必需的，但是添加的量过高也会显著抑制芽条的生根。 [0033] From Table 1, in the six kinds of rooting medium to induce buds, rooting is best to add a lmg / LIBA of 1 / 2MS medium buds on this medium for 30 days, it Rooting was 16.10%, the average number of roots per shoot bar for 2.58, with an average root length 2.51cm; buds hormone-free medium in the medium (MS and 1 / 2MS) and add three different auxin (l / 2MS + lmg / LIBA + lmg / LIAA + 2mg / LNAA) are not produce adventitious roots, indicating induction of buds auxin rooting may be necessary, but the amount added is too high will also significantly inhibited buds rooting . 在实验过程中还发现，采用传统方法诱导麻疯树再生不定芽芽条生根后所获得完整植株上的叶片容易发黄甚至脱落。 During the experiment also found that the use of traditional methods to induce regeneration of Jatropha adventitious buds after leaf buds on the roots of the plants are easy to obtain a complete yellow or even fall off. 总之，采用传统方法诱导麻疯树再生不定芽芽条生根的效果不佳，存在生根率较低，生根质量较差，难以获得生长状态良好的再生植株。 In short, using the traditional method of inducing poor Jatropha regeneration of adventitious buds buds rooting effect, there is a lower rate of rooting, rooting poor quality, it is difficult to obtain regenerated plants grew well. [0034] 实施例2 [0034] Example 2

[0035] 采用在培养基中添加L-谷氨酰胺的方法来促进麻疯树再生不定芽芽条生根的效果 [0035] The L- glutamine added to the medium in ways to promote Jatropha regeneration of adventitious buds buds rooting effect

[0036] S1.再生不定芽芽条的获取:从培养瓶中取出具有长度大于Icm且至少有2个伸展叶片的不定芽的外植体，用灭过菌的手术刀对伸长的再生不定芽进行横向切割(切割的方向与伸长的再生不定芽的中轴相垂直，切口要平齐)，切去其他部分，获取长度约为Icm的不定芽芽条。 . [0036] S1 adventitious buds bud regeneration of the acquisition: removed from culture flasks with a length greater than Icm adventitious buds and at least two extension leaves explants with a sterile scalpel to the elongation of the regeneration of adventitious lateral buds cutting (cutting direction and elongation of adventitious buds regeneration perpendicular axis, the incision should be flush), cut the other section for a length of about Icm adventitious buds buds.

[0037] S2.将SI获取的芽条以竖插方式(使芽条的中轴与培养基表面相垂直，芽条的形态学下端插入培养基中的深度为0.2~0.4cm。)接种至添加了0、4、8、16、32、64、128mg/LL-谷氨酰胺的生根培养基(大量元素减半的MS配方成分+30g/L蔗糖+100mg/L肌醇+0.3mg/LIBA+5g/L琼脂；pH5.8~6.0。)上培养，在10、20、30、40天时分别统计再生不定芽芽条生根的情况，所获得实验结果如表2、图2、图3和图4所示。 [0037] S2. The SI acquired bud bar to vertical interpolation method (so buds of the axis perpendicular to the surface of the medium, bud morphology of the lower end of the insertion depth of the medium is 0.2 ~ 0.4cm.) Inoculated Added 0,4,8,16,32,64,128mg / LL- glutamine rooting medium (MS macronutrients halved the recipe ingredients + 30g / L sucrose + 100mg / L Inositol + 0.3mg / LIBA + 5g / L agar;. pH5.8 ~ 6.0) on the train, at 10, 20 days, respectively, statistics regeneration of adventitious buds buds rooting the obtained results are shown in Table 2, Figure 2, Figure 3, and Figure 4.

[0038] 表2L-谷氨酰胺诱导麻疯树再生不定芽芽条生根的效果 [0038] Table 2L- Jatropha glutamine induced regeneration of adventitious buds buds rooting effect

[0039] [0039]

[0040] [0040]

[0041] 注:数据釆用SPSSStatisticsl7.0统计分析软件进行方差分析和邓肯多重比较(P兰0.05)，数据后字母不同表示处理间差异显著。 [0041] Note: The data preclude the use of statistical analysis software SPSSStatisticsl7.0 ANOVA and Duncan's multiple comparison (P lan 0.05), the difference between different letters represent data after treatment significantly. 生根率(％)=(再生出不定根的外植体数/总外植体数)X 100% ;平均每个外植体的根数(条)=再生不定根总数/再生出不定根的外植体数；平均根长(cm) =再生不定根长度的平均值。 Rooting rate (%) = (regeneration of adventitious root explants / total number of explants) X 100%; the average root number (bar) = each explant regeneration of adventitious roots total / regeneration of adventitious root explants count; average root length (cm) = average of adventitious root regeneration length.

[0042] 从表2可知，在未添加L-谷氨酰胺的生根培养基中，诱导芽条生根的效果较差，培养40天时生根率为最高，为22.76% ;而在添加L-谷氨酰胺的生根培养基中，随着添加的L-谷氨酰胺的浓度提高，芽条的生根率先增加，到16mg/L时，芽条的生根效果最佳，生根率最高为51.72%，平均每个芽条上的根数最多为6.61条，平均根长最长为5.36cm，继续提高L-谷氨酰胺的浓度，芽条的生根效果变差，生根率降低，到128mg/L时，生根率最高仅为14.81%，平均每个芽条上的根数最多为2.11条，平均根长最长为2.31cm。 [0042] From Table 2, without adding L- glutamine rooting medium to induce buds poor rooting effect for 40 days rooting rate was the highest 22.76%; while adding L- glutamine amide rooting medium, with the addition of L- glutamine concentration increased rooting buds lead increased to 16mg / L, the rooting of the best buds, rooting rate up to 51.72%, with an average per the number of buds bar up to 6.61, with an average root length up to 5.36cm, continue to increase the concentration of L- glutamine, rooting buds of the poor, rooting rate decreased to 128mg / L, the rooting the highest rate of only 14.81%, the average number of roots per shoot bar up to 2.11, with an average root length up to 2.31cm. 总之，在培养基中添加L-谷氨酰胺的浓度太低或者太高都将不利于芽条的不定根再生，适宜的浓度为16mg/l。 In short, in the medium supplemented L- glutamine concentration is too low or too high will not conducive buds adventitious root regeneration, suitable concentration of 16mg / l.

[0043] 从表2还可知，培养时间为10天时，芽条在添加了8~32mg/LL-谷氨酰胺的生根培养基中的生根率就已经达23.52~30.93%，尤其是在添加了16mg/LL_谷氨酰胺的生根培养基中芽条的生根效果最佳，生根率为最大值，为30.93%，平均每个芽条上的根数为2.25条，平均根长为1.36cm ;而此时芽条在未添加的L-谷氨酰胺的生根培养基中的生根效果不佳，生根率仅为8.69%，平均每个芽条上的根数为1.33条，平均根长为0.67cm。 [0043] From Table 2 also shows that the incubation time is 10 days, the buds in added 8 ~ 32mg / LL- glutamine rooting medium rooting rate has reached 23.52 ~ 30.93%, especially with the addition of 16mg / LL_ glutamine rooting medium rooting best buds, rooting rate maximum is 30.93%, the average number of roots per shoot strip is 2.25, with an average root length 1.36cm; At a time when the poor buds without adding L- glutamine rooting medium of rooting, rooting rate was 8.69%, the average number of roots per shoot bar for 1.33, 0.67 average root length cm. 随着培养时间的延长，芽条的生根率也是增加的，虽然培养40天时芽条的生根率均达最大值，但是培养30天时芽条的生根率与40天时的生根率没有显著差异，因此培养时间以30天为宜。 With the culture time, rooting rate buds also increased, although for 40 days buds rooting rate reached a maximum, but for 30 days buds rooting rate was not significantly different from the rate of 40 days of rooting, so incubation time to 30 days is appropriate.

[0044] 另外，对于那些没有生根的芽条，用灭过菌的手术刀对芽条进行横向切割(切割的方向与芽条的中轴相垂直，切口要平齐)，切去其形态学下端一部分(0.1~0.2cm)，将其重新接种至添加了16mg/LL-谷氨酰胺的生根培养基上进行再次生根培养，这样可以提高芽条的利用效率，增加完整的再生植株的数量。 [0044] In addition, for those who do not rooting buds with a sterile scalpel for cutting lateral buds (buds cutting direction of the axis perpendicular to the incision should be flush), cut its morphology the lower part (0.1 ~ 0.2cm), will be re-inoculated to add the 16mg / LL- glutamine were rooting rooting medium again, this can increase the utilization efficiency of buds, increasing the number of complete plant regeneration.

[0045] 从实施例1和实施例2可知:采用传统方法对麻疯树再生不定芽芽条进行不定根诱导时，对芽条的质量要求较高，一般是要求芽条的长度至少达到2cm，且至少带有4个叶片，这在一定程度上减少了可以用于进行生根培养的芽条的数量，因而会减少获得完整再生植株的数量；而采用在生根培养基中添加适宜浓度的L-谷氨酰胺的方法可以促使质量较差(如芽条长度约为1cm，只带有2~3个叶片)的芽条生根，这在一定程度上提高了可以用于生根培养的再生不定芽芽条的利用效率，最终可以增加获得的完整再生植株的数量，如果将此方法应用于麻疯树的遗传转化，将可能会提高获得完整转化苗的效率。 [0045] from Example 1 and Example 2 shows: the traditional method of Jatropha regeneration of adventitious buds buds were induced when adventitious roots on the bud of the high quality requirements, generally require at least the length of the shoot 2cm, and with at least four blades, which reduces the number may be used for rooting of shoots strip to some extent, and thus will reduce the number of plantlets obtained; and the use of appropriate concentrations added in rooting medium L- Glutamine ways to promote quality is poor (eg buds length of about 1cm, with only 2 to 3 leaf) shoots rooting section, which improves to some extent, can be used for regeneration of adventitious rooting Buds Article utilization efficiency, and ultimately increase the number of plantlets obtained, if this method is applied Jatropha genetic transformation, will likely raise seedlings for complete conversion efficiency. 同时，在实验过程中可以发现，添加了适宜浓度L-谷氨酰胺的实验组中的芽条培养30天后，不仅生根效果良好，所获得再生植株的生长状态也良好，叶片鲜绿，不易发黄。 Meanwhile, during the experiment can be found in the experimental group added in the appropriate concentration of L- glutamine buds cultured for 30 days, not only rooting good effect, obtained regenerated plants growing state is also good, leaf-green, easy hair yellow.

[0046] 众所周知，诱导再生不定芽生根对于建立与完善一个良好的再生体系有着至关重要的意义，因为如果仅仅可以获得再生不定芽，却不能诱导其生根，将不能获得独立生活的植株。 [0046] It is well known to induce regeneration of adventitious rooting is essential for the establishment and improvement of the regeneration system has a good sense, because if you can only get regenerated adventitious buds, but can not induce its roots, the plants will not get to live independently.

[0047] 综上所述，由表1和表2中的数据可知，麻疯树再生不定芽芽条在添加16mg/LL谷氨酰胺的培养基中的生根效果显`著优于采用传统方法的生根效果，同时在较短时间内(10天)就可以达到较好的生根效果(生根率最高为30.93%)，最后还可以获得生长状态也良好的完整再生植株。 [0047] In summary, we can see from the data in Table 2 and Table 1, jatropha regeneration of adventitious buds buds adding 16mg / LL glutamine rooting medium effect was `significantly better than traditional methods rooting effect, but in a relatively short period of time (10 days) you can achieve better rooting effect (rooting rate up to 30.93%), and finally get a good growth state also complete plant regeneration.