苹果生长素响应因子MdARF5的克隆与功能鉴定

【目的】分离苹果生长素响应因子MdARF5（Auxin Response Factor 5）,分析其对生长素的响应,鉴定其在调节花青苷合成过程中的作用,揭示MdARF5的生物学功能,为进一步研究生长素对花青苷的调节提供理论依据。【方法】以‘嘎拉’苹果（Malus×domestica ‘Royal Gala’）为材料,利用同源克隆技术,克隆得到一个ARF（Auxin Response Factor）转录因子,并将其命名为MdARF5。利用MEGA5.0软件构建多物种间系统进化树。通过农杆菌介导的遗传转化获得转基因苹果愈伤组织。比较野生型和转基因苹果愈伤组织花青苷积累的差异。利用烟草叶片瞬时转化试验,分析MdARF5对MdMYB1的转录调控。【结果】克隆获得苹果生长素响应因子MdARF5（序列号：MDP0000143749）,该基因CDS为2 691 bp,编码含有896个氨基酸的蛋白。系统进化树分析表明,苹果MdARF5与梨PbARF5同源性最高。基因表达分析显示,该基因响应生长素处理,并且与花青苷合成相关基因表现出相反的表达模式。在苹果愈伤组织中超表达MdARF5,其花青苷积累较野生型显著降低,表明MdARF5在调控花青苷积累过程中发挥重要作用。对苹果MdMYB1启动子序列进行分析,发现其序列包含一个MdARF5的结合位点。烟草瞬时表达试验显示,MdARF5能够抑制MdMYB1的表达。【结论】推测苹果MdARF5可能通过直接抑制MdMYB1的表达负调节花青苷的积累。     

金边黄杨不同生育期内源激素含量差异分析

为了分析和探讨金边黄杨不同生育期顶端分生组织及叶原基内源细胞分裂素(ZRs、DHZRs、iPAs)及内源生长素(IAA)的含量差异,利用植物激素ELISA方法对金边黄杨的顶端分生组织和不同大小茎尖的内源激素进行了测定。结果表明,金边黄杨的顶端分生组织的内源ZRs、DHZRs、iPAs、IAA及CTK/IAA值在不同的生育期均无显著差异;但在相同生育期时,金边黄杨茎尖的内源ZRs、DHZRs、iPAs、IAA及CTK/IAA值会随着保留的叶原基个数的增多而呈下降的趋势。这说明了金边黄杨顶端分生组织内源激素不会随着其生长而改变,并且间接地证明了顶端分生组织周围叶原基的内源激素含量与顶端分生组织不一致,它的存在会影响测定顶端分生组织内源激素含量的准确性。     

水稻矮缩病毒对3种内源激素含量及代谢相关基因转录水平的影响

本文以抗病品种宜香2292(Oryzas ativa L. ssp. indica cv. Yixiang 2292)为材料,采用高效液相色谱技术(HPLC)和Real-time PCR技术研究了水稻矮缩病毒(Rice dwarfvirus,RDV)胁迫下水稻内源赤霉素(GA3)、生长素(IAA)和脱落酸(ABA)的动态变化。结果表明,受RDV侵染后,病株体内GA3含量显著低于健株,在显症后的第1d和第10d最为明显,分别较健株低6.28和5.92倍;IAA含量呈现波动变化,但病株体内的IAA含量始终较健株低,在显症后的10d最为明显,比健株低3.58倍;与GA3和IAA相反,病株体内ABA的含量始终高于健株,显症后的第1d和第13d最为明显,分别较健株高2.29和2.84倍。Real-time PCR定量检测了植物内源激素相关基因mRNA的表达,结果显示,GA3代谢相关的氧化还原酶基因表现为下调,而IAA和ABA代谢相关的Cullin-1和P-glycoprotein1基因表现出不同程度的上调。以上结果表明:水稻矮缩病的症状表现可能与病株体内的植物内源激素失调有关。     

啤酒花组培苗生根移栽培养试验研究

摘要：本试验采用第六代啤酒花试管苗为材料,研究了不同浓度生长素、培养基营养元素及碳源葡萄糖浓度对试管苗生根移栽的影响。结果表明IBA浓度为0.05mg/L是生根效果最佳;无根苗在培养基B5(大量)+MS(微量、有机物)中,组织充实,抑制徒长;当培养基中糖浓度为15%,苗木的生根率及移栽成活率最高。     

Vegetative propagation of twelve fodder tree species indigenous to the Sahel,West Africa

In the Sahel region, many woody species are used as fodder because of their high nitrogen content, especially during the dry season, in contrast to grasses and crop residues. Unfortunately, this resource is being threatened by regular pruning, increasing livestock browsing and impeding their natural regeneration. Therefore, there is a need to find appropriate regeneration options to sustain fodder production. Thus, a series of vegetative propagation experiments were conducted to identify the most appropriate multiplication methods of some of the species used as fodder. This series included testing the effects of naphthalene acetic acid (NAA) and indole butyric acid (IBA) on rooting ability of stem cuttings of 12 species: Afzelia africana, Balanites aegyptiaca, Bauhinia rufescens, Commiphora africana, Faidherbia albida, Ficus gnaphalocarpa, Guiera senegalensis, Kigelia africana, Pterocarpus erinaceus, Pterocarpus lucens, Pterocarpus santalinoides and Terminalia avicennioides. The series also evaluated the impacts of the size (C. africana) and the age (P. erinaceus) of the cuttings on their rooting success. Pterocarpus santalinoides was revealed to be an easy-to-root species, showing a high percentage of rooted cuttings (up to 88±3%), followed by P. erinaceus, P. lucens and B. aegyptiaca (up to 37±4%, 31±5% and 26±6%, respectively), in contrast to A. africana, cuttings of which did not root at all. The application of NAA and IBA enhanced the number of roots and secondary roots per rooted cutting. Large-diameter cuttings (15–20?mm) of deeply planted C. africana gave the highest percentage of rooted cuttings (78.3±4.4%). Tree pruning improved the rooting capacity of cuttings collected from P. erinaceus with one-month resprouts giving the highest percentage of rooted cuttings (50.0±6.7%). These results indicate that P. santalinoides, C. africana, P. erinaceus, P. lucens and B. aegyptiaca are amenable to vegetative propagation as an alternative to sexual propagation.     

缺磷胁迫下番茄侧根形成与miR164及NAC1表达的关系

为了研究番茄幼苗在缺磷胁迫下根系形态发育与生长素、生长素信号转导途径中的转录因子NAC1,以及调控NAC1 表达的上游miR164之间的关系。试验以5和500 μmol/L磷浓度作为缺磷胁迫和对照,检测了外源生长素NAA（1-naphthalene acetic acid）及生长素抑制剂NPA（N-1-naphthylphthalamic acid）对侧根形成的影响; 同时采用RT-PCR检测了NAC1和miR164在缺磷胁迫下的时序表达。结果表明,缺磷胁迫下侧根大量形成与生长素及其运输密切相关,在侧根迅速形成的24 h内,NAC1的表达在缺磷胁迫下增强; 而其上游的miR164表达降低,从而揭示了缺磷胁迫下侧根形成与miR164调节NAC1表达之间的关系。     

Perspectives of Using L-Tryptophan for Improving Productivity of Agricultural Crops: A Review

Plant growth regulators are biologically active signaling molecules that regulate a number of plant physiological processes. Auxin (indole-3-acetic acid) is an important plant growth regulator and is synthesized within plant tissues through L-tryptophan (L-TRP)-dependent and -independent pathways. It has been found that plants respond to exogenously applied L-TRP due to insufficient endogenous auxin biosynthesis. The exogenous application of L-TRP is highly significant for normal plant growth and development. L-tryptophan is applied through foliar spray, seed priming, and soil application. Soil-applied L-TRP is either directly taken up by plants or metabolized to auxin by soil microbiota and then absorbed by plant roots. Similarly, foliar spray and seed priming with L-TRP stimulates auxin synthesis within plants and improves the growth and productivity of agricultural crops. Furthermore, L-TRP contains approximately 14% nitrogen (N) in its composition, which is released upon its metabolism within a plant or in the rhizosphere and plays a role in enhancing crop productivity. This review deals with assessing crop responses under the exogenous application of L-TRP in normal and stressed environments, mode of action of L-TRP, advantages of using L-TRP over other auxin precursors, and role of the simultaneous use of L-TRP and auxin-producing microbes in improving the productivity of agricultural crops. To the best of our knowledge, this is the first review reporting the importance of the use of L-TRP in agriculture.     

对生和互生玉米内源激素含量的差异分析

利用ELISA方法测定近等基因系玉米(Zeamays)对生、互生玉米的不同组织和不同生育期内源细胞分裂素ZRs(zeatin riboside group)、DHZRs(dehydro zeatin riboside group)、iPAs(isopentenyladenosine group)以及内源生长素IAA(indole acetic acid)含量差异。结果表明,不同叶序玉米在不同生育期幼胚、成熟花粉和成熟叶片的内源ZRs、DHZRs、iPAs和IAA均无显著差异;不同生育期对生叶序玉米幼苗顶端分生组织和成熟苗顶端雄幼穗ZRs、DHZRs和iPAs含量显著高于同期近等基因系的互生玉米,IAA含量在两者之间无显著差异;对生叶序玉米幼苗顶端分生组织、成熟苗顶端雄幼穗CTK/IAA(CTK表示ZRs、DHZRs和iPAs含量相加)比值显著高于同期近等基因系互生叶序玉米,但在不同生育期幼胚、叶片及花粉内均无显著差异。对生叶序的形成与生长点组织细胞分裂素含量增加密切相关。     

IAA对水稻根毛形成与水通道蛋白基因表达关系的研究

【目的】探讨气培条件下,IAA极性运输对水稻根毛形成及水通道蛋白基因表达的影响。【方法】以超表达OsPIN1a的转基因水稻和野生型水稻为研究材料,当种子根长为0.5-1.0 cm时,分别用不同浓度IAA、IAA和生长素极性运输输出载体抑制剂、IAA和生长素输入载体抑制剂复合处理的琼脂块贴在根尖一侧,黑暗条件下培养12 h后观察根毛生长情况,测定根毛的密度、长度及根毛形成后根尖部分相对含水量,并用激光共聚焦显微镜观察根尖及根毛内的OsPIN1a-GFP亚细胞定位,再通过半定量RT-PCR检测根毛形成前后水通道蛋白基因表达。【结果】（1）在0-5.0 mg·L^-1 IAA浓度范围内,随IAA浓度增加,根毛长度和密度也增加,当浓度超过5.0 mg·L^-1时,根毛长度和密度不再增加,根的生长受到显著抑制,2.5 mg·L^-1 IAA诱导效果最好;（2）只有根尖分生区IAA处理后,才能诱导新的根毛区形成根毛,而其他区域处理后无法诱导根毛形成;（3）根毛形成不受低浓度的生长素极性运输输出载体抑制剂TIBA、NPA和IAA复合处理的影响,高浓度TIBA、NPA和IAA复合处理则显著抑制根毛形成及根的生长;（4）低浓度生长素极性运输输入载体抑制剂CHPAA和IAA复合处理能显著抑制根毛形成和根的生长;（5）IAA不但能诱导根毛形成,也能诱导多种水通道蛋白基因表达,提高根尖相对含水量;（6）根尖及根毛内的OsPIN1a基因的表达和OsPIN1a-GFP含量均受IAA诱导,并在根毛细胞内大量表达。【结论】在气培条件下,根毛形成需要IAA诱导,而IAA诱导根毛形成过程需要生长素极性运输输入和输出载体蛋白的参与,其中输入过程对根毛的形成影响最大。IAA极性运输输入或输出载体蛋白特异性抑制剂处理显著降低根毛形成和根的生长,因此OsPIN1a在根毛形成中起着重要作用,同时根毛水通道蛋白基因表达量增加,提高了根尖相对含水量,减缓了气培条件下水稻根尖的水分胁迫。