Genomic analysis of isopentenyltransferase genes and functional characterization of TaIPT8 indicates positive effects of cytokinins on drought tolerance in wheat

Cytokinins(CKs)function in plant development and during stress responses,but their role in drought tolerance in wheat is unknown.In the present study,24 isopentenyltransferase(IPT)genes,encoding ratelimiting enzymes in CK biosynthesis were identified in the wheat genome.The chromosomal locations and structures of the genes,protein properties,and phylogenetic relationships were characterized.ATP/ADP TaIPT genes showed tissue-specific expression.TaIPT2,TaIPT7,and TaIPT8 expression was rapidly induced by 0.5–1 h drought treatments,which decreased to low levels after 2 h drought treatment,as did most other TaIPT genes.TaIPT8-5a/5b/5d triple mutants showed decreased levels of tZtype CK under normal and drought conditions and reduced drought tolerance,which,however,did not manifest as phenotype alterations.By contrast,transgenic wheat plants with drought-induced TaIPT8showed increased drought tolerance.Our study provides a foundation for further investigation of TaIPT genes and novel insights into the role of CKs in the drought response of wheat.

白菜型油菜IPT基因家族鉴定及表达分析

异戊烯基转移酶(isopentenyltransferase,IPT)是细胞分裂素生物合成的第一限速酶,为分析IPT基因家族在白菜型油菜生长中的功能,利用生物信息学方法从其基因组中鉴定出13个BrIPT基因,它们不均匀地分布在7条染色体上。BrIPTs可分为4个亚族,各家族成员均含有8~10个保守基序和1~2个UTR区。BrIPT基因的启动子区域包含众多响应元件。BrIPT基因家族成员受环境因子、生物激素及防御与应激调控。qRTPCR结果显示,tRNA-IPT基因BrIPT4、BrIPT6、BrIPT9在白菜型油菜体内各个部位均有表达。相比于苗期,成熟期白菜型油菜IPT基因的表达量更高,为后续深入研究IPT基因家族成员的生理生化功能提供了理论依据。

细胞分裂素受体基因SlHK4突变对番茄开花时间的影响

[目的]本研究旨在解析细胞分裂素受体基因SlHK4突变对番茄开花时间的影响,为揭示细胞分裂素途径调控植物开花的分子机制奠定基础。[方法]以细胞分裂素受体基因SlHK4的2个纯合突变株系slhk4-4和slhk4-5及野生型(WT)番茄为材料,统计各株系开花时间,并在发芽26 d后取植株茎尖进行石蜡切片显微观察、响应细胞分裂素的荧光信号检测以及转录组测序等分析。[结果]slhk4-4和slhk4-5突变体的始花节位与WT相同,但始花时间分别比WT晚6和4 d。与WT相比,slhk4-5茎尖花分生组织分化推迟,细胞显著减小,细胞数目略少;slhk4-4与细胞分裂素响应启动子TCSn驱动的绿色荧光蛋白基因GFP过表达株系的杂交F 1代茎尖中,响应细胞分裂素的GFP荧光信号显著减弱;slhk4-5茎尖中细胞周期正调控基因Cyclin-B1-2表达下调,开花正调控基因FUL2、SOC1表达上调,而花发育基因AGL12和AGL19表达下调,开花调控相关基因的表达变化趋势与slhk4-5开花推迟的性状变化不完全一致。差异表达基因(DEG)的功能分析显示,57.6%具有功能注释的DEG是富集于光合作用、呼吸作用、蛋白合成和代谢等相关通路中的结构基因,其中88.2%在slhk4-5茎尖中的表达下调,表明slhk4茎尖的细胞活性很可能弱于WT。[结论]细胞分裂素受体SlHK4的功能缺失导致slhk4茎尖细胞分裂素信号减弱,花分生组织细胞减小,活性降低,分化延后,进而导致slhk4开花推迟。SlHK4是番茄开花时间的正调控因子。

生长素和细胞分裂素诱导植物胚性愈伤组织研究进展

植物胚性愈伤组织在植物研究与应用中是一种重要的试材,有很多关于植物胚性愈伤组织诱导方法研究的报道。生长素和细胞分裂素被广泛应用于胚性愈伤组织的诱导,是胚性愈伤组织诱导的关键因子。阐述了生长素和细胞分裂素在植物胚性愈伤组织诱导及体细胞胚胎发生过程中的作用,讨论了其在植物胚性愈伤组织诱导中的作用、使用方法与原则,为愈伤组织诱导、愈伤组织胚性化诱导等研究中植物生长调节剂的使用提供依据。

细胞分裂素信号调控植物雌配子体发育的研究进展

雌配子体发育是被子植物有性生殖过程的重要阶段,它的顺利进行是保证植物完成整个生命周期的前提。细胞分裂素是植物自身合成的一类重要激素,广泛参与植物生长发育等一系列生命过程。在综述细胞分裂素代谢、信号感知和转导的基础上,重点介绍细胞分裂素信号在植物雌配子体发育过程中的功能,旨在从细胞分裂素角度揭示植物雌性生殖发育机理,为提高农作物产量及其遗传改良研究提供理论依据。

超高效液相色谱-三重四级杆质谱联用法同时测定荔枝中4种细胞分裂素

【目的】应用超高效液相色谱-三重四级杆质谱联用仪(UPLC-MS/MS),建立一种同时测定荔枝反式玉米素核苷(tZR)、异戊烯基腺嘌呤核苷(IPR)、二氢玉米素(DHZ)、异戊烯基腺嘌呤(IP)4种细胞分裂素含量的方法。【方法】0.25 g荔枝样品用乙腈-水(80∶20,体积比)溶液浸提8 h,经Bond Elut Plexa PCX固相萃取柱纯化,2.5%氨水甲醇洗脱后过0.22μm有机滤膜检测。选取XSelect HSS T3色谱柱,以甲醇和5 mmol·L^(-1)甲酸铵水溶液为流动相进行7 min梯度洗脱,采用电喷雾正离子(ESI+)模式电离,选择反应监测(MRM)模式对细胞分裂素进行定量。【结果】4种细胞分裂素的检出限和定量限分别低于18.12和60.39 pg·g^(-1),在0.05~50 ng·mL^(-1)质量浓度范围内呈良好的线性关系,线性相关系数(r2)大于0.999。在高、中、低浓度三种加标水平下,4种细胞分裂素的平均回收率为80.0%~108.2%,标准偏差为0.8%~15.5%。采用建立的方法可以同时检测出荔枝果皮、果肉、种子、幼果、叶片和果柄离区中4种细胞分裂素含量。【结论】该方法简便、快速、灵敏、准确,适用于荔枝不同组织中细胞分裂素含量的测定。

PbGIF1 promoting cell-proliferation in pear fruit is transcriptionally activated by Pb RR1

As a cell proliferation regulator involved in wide biological processes in plants,GRF-INTERACTING FACTOR(GIF)controls different tissues development.However,whether GIF participates in fruit development remains unclear.According to transcriptome data,we identified PbGIF1was highly expressed during fruit development in cytokinins induced parthenocarpy pear.In the present study,the biofunction of PbGIF1 was initially verified.Overexpression of PbGIF1 promoted fruit size of transgenic tomato.The size of flesh fruit was not affected by cell expansion but the cell proliferation was promoted by overexpressing Pb GIF1.The accelerated cell proliferation process was also observed in PbGIF1-overexpressed transgenic pear fruit calli.The transcriptional regulation of cytokinins on PbGIF1 was further confirmed by exogenous CPPU treatments in pear fruitlets.To investigate the underlying mechanism,the cytokinins-responded factor,PbRR1,was further focused on.The results of Yeast-one-hybrid assay suggested that PbRR1 can bind to the promoter sequence of PbGIF1.The transcriptional activation of PbRR1 on PbGIF1 was also confirmed by Dual-Luciferase assays.Taken together,the results showed that cytokinins control pear fruit development via the transcriptional activation of PbGIF1 by PbRR1.

EXOGENOUS Ca^(2+) INHIBITS SENESCENCE-RETARDING EFFECT OF CYTOKININS IN DETACHED RICE LEAVES

Cytokinins and Ca 2+ singly retarded senescence of detached rice leaves.When Ca 2+ was applied together with cytokinins,the effectiveness of cytokinins was significantly reduced.Ca 2+ and cytokinins did not stimulate ethylene production synergistically,ruling out the possibility that ethylene was involved in the inhibition of cytokinin induced senescence retarding effect by Ca 2+ .The experiment with specific compounds known to increase (Ca ionophore A23187),or decrease (EGTA,LaCl 3,Verapamil,chlorpromazine) cytosolic Ca 2+ level indicated that the elevated cytosolic Ca 2+ retards senescence.

Improving dryland maize productivity and water efficiency with heterotrophic ammonia-oxidizing bacteria via nitrification and cytokinin activity

A two-year field experiment conducted under dryland conditions in semi-humid and drought-prone regions of China aimed to assess the effect of ammonia-oxidizing bacterial on maize water use efficiency and yield.A heterotrophic ammonia-oxidizing bacteria(HAOB)strain S2_8_1 was used.Six treatments were applied:(1)no irrigation+HAOB strain(DI),(2)no irrigation+blank culture medium(DM),(3)no irrigation control(DCK),(4)irrigation+HAOB(WI),(5)irrigation+blank culture medium(WM),and(6)irrigation control(WCK).Results revealed that HAOB treatment increased maize growth,yield,and water use efficiency over controls,regardless of whether the year was wet or dry.This improvement was attributed to the accelerated nitrification in the rhizosphere soil due to HAOB inoculation,which subsequently led to increased levels of leaf cytokinins.Overall,these findings suggest that HAOB inoculation holds promise as a strategy to boost water use efficiency and maize productivity in dryland agriculture.

苜蓿链霉菌TX21抑菌作用确定及促生物质的鉴定

链霉菌是一种重要的微生物资源,具有广泛的应用价值。本研究从土壤中筛选出一株对多种病原菌有抑制效果的菌株,编号TX21,其中对玉蜀黍平脐蠕孢的抑菌效果最优,为50.0%。采用浸种及喷施的方法探究了菌株TX21发酵上清液对水稻种子和黄瓜幼苗的促生效果,结果显示发酵上清液能使水稻种子胚根和胚芽分别增长48.5%和47.1%,黄瓜幼苗根系和鲜重分别增加了37.6%和47.0%,表现出良好的促生作用;通过形态学观察、生理生化和16S r RNA分子生物学鉴定苜蓿链霉菌Streptomyces alfalfae。利用黄瓜子叶叶绿素试验及UHPLC-MS方法明确了发酵上清液的有效促生物质,证实其中含有2-甲硫顺式玉米素核苷(ms^(2)io^(6)A,分子式:C_(16)H_(23)N_(5)O_(5)S)。

5种不同的植物激素对白花前胡种子萌发的影响

目的 研究生长素(IAA)、细胞分裂素(6-BA)、赤霉素(GA3)、脱落酸(ABA)和乙烯(ET)5种植物激素对白花前胡种子萌发的影响。方法 以宁国产并在室温放置12个月的白花前胡种子为材料,把5种不同的植物激素均配制成4个梯度浓度(1×10^(-1)、1×10^(-2)、1×10^(-3)、1×10^(-4)mg·mL^(-1)),对白花前胡种子进行浸种处理。通过培养皿萌发试验,分析种子的发芽率、根长和鲜重的变化情况。结果 在4个梯度范围内,6-BA对白花前胡种子的生长状态整体起抑制作用。把1×10^(-1)mg·mL^(-1)和1×10^(-2)mg·mL^(-1)设置为高浓度,1×10^(-3)mg·mL^(-1)和1×10^(-4)mg·mL^(-1)设置为低浓度。其中6-BA无论是高浓度还是低浓度均对白花前胡种子的生长状态整体起抑制作用。而GA3和ET在高浓度(1×10^(-1)mg·mL^(-1)和 1×10^(-2)mg·mL^(-1))及ABA和IAA在低浓度(1×10^(-3)mg·mL^(-1)和1×10^(-4)mg·mL^(-1))下均对白花前胡种子的萌发率、根长和鲜重都表达出明显的促进作用。结论 植物激素对白花前胡种子的生长发育均有不同程度的影响,其中ET在1×10^(-2)mg·mL^(-1)时,对白花前胡种子的生长发育影响最为显著,白花前胡种子的萌发率,最大根长和最大鲜重均高于其他组。

4种化学调控剂对上部烟叶开片及产质量的影响

以烤烟品种鲁烟1号为材料,在胶州进行大田试验,研究打顶后喷施4种化学调控剂(海藻素、赤霉芸苔素、细胞分裂素和光碳核肥)对上部烟叶开片和产质量的影响。结果表明:4种化学调控剂对调控烟株生长及改善烟叶品质均有一定作用。其中,细胞分裂素最能显著促进上部烟叶长、叶宽和叶面积的增加,促进上部叶开片,提高烟叶产量、产值、均价和上等烟及上中等烟比例等经济性状指标,改善烤后上部烟叶物理性状和化学成分协调性,提高烟叶品质。

‘小果甜柿’组织培养中的抗褐化研究

为降低‘小果甜柿’组织培养中的褐化现象,以其组培继代苗为试材,从基本培养基类型、细胞分裂素种类浓度及抗褐化剂种类浓度三个方面,对‘小果甜柿’进行了组织培养中的抗褐化研究。试验结果表明,相对于(1/2N)MS、(3/4N)MS和MS基本培养基,DKW培养基褐化最轻,褐化仅限于植株基部,未扩散至整个培养基而引起严重褐化;单独使用细胞分裂素ZT抗褐化效果较好,而单独使用6-BA或ZT+6-BA组合,褐化程度均较重;抗褐化剂AC和核黄素对‘小果甜柿’组培继代苗褐化有抑制效果,PVP对褐化无抑制效果,AC优于核黄素和PVP,但AC和核黄素会抑制植株生长而影响继代增殖。因此,综合而言,以DKW为基本培养基,添加细胞分裂素ZT(2.0mg/L),不添加抗褐化剂时,‘小果甜柿’组培继代苗增殖及抗褐化效果表现最佳。

Seasonal changes of endogenous ABA and cytokinins in environmental adaptation of different ecotypes of reed plants

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Review: Plant Growth Hormone Cytokinins Control the Crop Seed Yield

This review chronicles the development of the cytokinin research during the last 30 years. Cytokinin and auxin are the two major plant growth hormones that control virtually all aspects of growth and development in higher plants. The pathways for cytokinin biosynthesis and metabolism have been characterized by the identification of isopentenyl pyrophosphate transferase, cytokinin oxidases, cytokinin hydroxylase, zeatin cis-/trans-isomerase, cytokinin phosphoribosyl hydrolases, cytokinin-specific riboside phosphorylase, and others enzymes. Loss-of function mutant phenotypes of cytokinin degradation/activating enzymes indicate the regulation of concentration and spatial distribution of bio-active cytokinin plays a pivotal role in the increase in panicle size, in the numbers of floral organs, and eventually in seed yield. One of the most fundamental questions in the cytokinin field is one concerning the prevalence of cis-zeatin in monocotyledonous crops (rice and maize) and in dicotyledonous legumes (pea, chickpea) and potato/sweet potato. A hypothesis is that cis-zeatin is synthesized by the cis-specific hydroxylation of the terminal methyl group of N6-isopentenyl side chain of N6-isopentenyl adenosine (i6Ado) or of their mono-, di-, or tri-phosphates catalyzed by the cis-specific hydroxylase. A second potential pathway is the isomerization of trans-zeatin to cis-zeatin by zeatin cis-/trans-isomerase. A second fundamental question to be addressed is the physiological role of cis-zeatin. Some have argued for a special function of cis-zeatin to account for the prevalence of the cis-zeatin in the plant kingdom from algae to higher plants.

Effect of Macronutrients, Cytokinins and Auxins, on <i>in Vitro</i>Organogenesis of <i>Thymus vulgaris</i>L.

The present study reports an efficient protocol for in vitro propagation of Thymus vulgaris L., an aromatic and medicinal plant in Morocco. Initially, we performed in vitro multiplication of Thymus vulgaris explants existing in the laboratory and obtained from micropropagation by shoot tip culture. Afterwards, we have evaluated the effect of six macronutrients. After that, seven cytokinins (Kin, BAP, 2iP, DPU, Adenine, Zeatine and TDZ) in three different concentrations (0.46, 0.93, 2.32 μM) have been evaluated to optimize cultures multiplication and elongation. Moreover, the effect of three auxins (IAA, IBA and NAA) at 0.57 μM, combined to 4 cytokinins (Kin, BAP, DPU and Ad.) at 0.46 μM, on shoot rooting has been studied. Thereby, MS medium has been proved the most favorable for plantlets growing. Also, we found that the addition of certain cytokinins, specifically 0.46 Kin, 0.46 and 0.93 BAP, 0.46 2iP, 0.46 DPU, 0.46 Ad. and 0.46 Zeat., ensures better multiplication and growth of vitroplants. In addition, multiplication and rooting of cultures were well optimized after addition 0.46 Kin + 0.57 IAA or NAA, 0.46 DPU + 0.57 IBA and 0.46 Ad. + 0.57 IBA combinations to the culture medium. Lastly, plantlets with roots were successfully acclimatized to ex-vitro conditions and these latter served as a source to establish in vitro culture again.

EFFECT OF CYTOKININS AND AUXINS ON THE GROWTH OF FREE-LIVING CONCHOCELIS OF PORPHYRA YEZOENSIS

1AA 3-Indolylacetic acid, NAA a-Naphthylacetic acid and cytokinins in PESI culture medium were used in a study on the effects of plant hormones on the growth of free-living conchocelis of Porphyra yezoensis which showed that its growth in medium with cytokinins, 1AA and NAA was more rapid than that in medium with non-phytohormones; that the optimal concentrations for promoting growth were 10μg/L for IAA and ZA (Zeatin), and 0.1 μg/L for BA 6-Benzyl amino purine and KIN 6-Furfurylamino- purine. Mix use of NAA, 1AA and cytokinins, NAA/ZA 1-1000/1μg/L, NAA/BA 10/1-1000 μg/L, NAA/KIN 1/1-1000 μg/L promoted growth. 1AA/ZA 0.1-1/0.1-1μg/L; 1AA/BA 0.1-1/0.1-10 μg/L IAA/KIN 1/0.1-1000μg/L also promoted growth.

Effect of Cytokinins on the Micropropagation of Carob(Ceratonia siliqua L.)through Shoot Tip Culture

In order to determine the most suitable cytokinin for the micropropagation of carob (Ceratonia siliqua L.), the effect of four cytokinins: BAP, zeatin, kinetin and 2-iP, was tested on explants derived from young seedlings of seven days. Organogenesis is better in the presence of BAP (0.5 mg/l), while buds growth is favored by zeatin (0.5 mg/l). The combination of the most suitable cytokinin (BAP at 0.5 mg/l) with IBA improves the caulogenesis when the concentration of this latter is low (0.1 mg/l);high concentrations of IBA have an inhibitory effect on elongation and neoformation of shoots and leaves. The multiplication and growth of shoots are more favorable on WPM medium in the presence of BAP (0.5 and 1 mg/l) alone or combined with 0.5 mg/l with GA3, while rooting is mainly favored by IBA, especially at 2 mg/l.

Polyamine Alterations in Isolated Zucchini Cotyledons Grown in Presence of Cytokinins and Cu²⁺

Polyamines are small nitrogen-containing organic molecules, which are widely distributed in plants. They are involved in the regulation of normal plant growth and developmental processes. In this study we examined the role of polyamines on the growth of Cucurbita pepo L.(zucchini) cotyledons incubated on solutions of different types of cytokinins (BA—N6-benzylaminopurine or 4PU-30—N1-(2-chloro-4-pyridyl)-N2-phenylurea) and copper in excess. We found that endogenous polyamines, and mainly the conjugated fraction, are involved in the cell division processes of isolated zucchini cotyledons and their changes are related to the specific action of the used growth regulating factors.

水稻种子胚形态与胚乳组成对稻米留胚特性的影响

【目的】稻米留胚能够提高稻米营养品质,但稻米留胚特性与胚性状、胚乳成分的关系等相关生物学机制并不明确。因此,有必要深入研究不同类型稻米品种留胚率差异及其影响因素。【方法】选取粳稻中软米型和普通型两类典型留胚材料及非留胚材料为试材,首先切片观察两类品种籽粒胚发育特点,继而对比表观直链淀粉含量(apparent amylose content,AAC)和离层果胶含量,最后以外源细胞分裂素(cytokinin,CTK)调控,比较胚性状、胚乳组成及留胚率差异。【结果】与非留胚材料相比,软米型和普通型留胚材料具有胚内幅和外幅较大,胚发育速度快,胚体积较大的特点;籽粒充实度好,粒形更宽、厚;同时离层果胶含量也较高。外施CTK后,软米型留胚材料中后期胚发育速度加快,普通型材料胚发育减缓;两类材料的AAC、果胶含量、胚比重及留胚率均提高。【结论】除胚性状对两类品种留胚率均有明显影响外,胚乳组成主要影响软米型材料留胚特性,而粒形是影响普通型材料留胚特性的重要因素。CTK则通过影响籽粒不同性状对不同类型材料留胚率进行调控。这为后续高留胚水稻品种的培育和栽培调控提供了理论依据和指导方向。