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Purification, immunological, and functional characterization of MxFIT in Malus xiaojinensis 小金苹果MxFIT基因的纯化、免疫学及功能研究
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-06-11 DOI: 10.32615/BP.2021.004
L. Yin, X. L. Chen, S. L. Ma, F. Li, F. Zhou, Yongyan Wang
MxFIT is a FER-like iron deficiency induced transcriptional factor in Malus xiaojinensis. Here, we described the heterologous expression of MxFIT in Escherichia coli BL21 (DE3) host cells. The E. coli harboring the recombinant construct pET-MxFIT was efficiently induced to express the MxFIT protein at a high level and the optimal profile for MxFIT expression was investigated. By inoculating a New Zealand rabbit with purified MxFIT-His fusion protein, a high specific anti-MxFIT antiserum was achieved. Western blot analysis showed MxFIT protein expression was induced in roots when iron supply was limiting and was inhibited when iron supply was excessive. In leaves, there was almost no expression irrespective of iron supply. The localization of MxFIT on transverse section of root indicated that MxFIT participated in iron deficiency response. Over-expression of MxFIT in transgenic tobacco suspension cells showed that MxFIT increased iron absorption under insufficient iron supply. This study provides a basis for further investigating the underlying mechanism of high iron absorption efficiency in M. xiaojinensis.
MxFIT是小金苹果铁缺乏的转录因子。在这里,我们描述了MxFIT在大肠杆菌BL21 (DE3)宿主细胞中的异源表达。我们高效地诱导携带重组构建物pET-MxFIT的大肠杆菌高水平表达MxFIT蛋白,并研究了MxFIT的最佳表达谱。用纯化的MxFIT-His融合蛋白接种新西兰兔,获得了高特异性的抗mxfit抗血清。Western blot分析显示,当铁供应不足时,MxFIT蛋白在根中被诱导表达,当铁供应过剩时,MxFIT蛋白在根中被抑制表达。在叶片中,与铁供应无关,几乎没有表达。MxFIT在根横切面的定位表明MxFIT参与了缺铁反应。转基因烟草悬浮细胞中过表达MxFIT表明,在铁供应不足的情况下,MxFIT增加了铁的吸收。本研究为进一步探讨小金草高铁吸收效率的潜在机制提供了基础。
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引用次数: 0
Lysigenous aerenchyma formation: responsiveness to waterlogging in oil palm roots 油棕根部溶解性通气组织的形成:对涝渍的响应
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-06-11 DOI: 10.32615/BP.2021.002
S. Nuanlaong, S. Wuthisuthimathavee, P. Suraninpong
Oil palm (Elaeis guineensis Jacq.) responds to waterlogging stress by induction of lysigenous aerenchyma tissues, which facilitates the flow of oxygen through their root tissues for survival under waterlogged conditions. Thus, the morphological and genetic adaptation involved in lysigenous aerenchyma formation in the roots of the oil palm genotype Deli × Calabar under waterlogging stress was evaluated. This study found the highest number of dead cells after waterlogging stress for 2 d in the secondary root, while the percentage of root porosity was increased with increasing of time in both roots, especially at 1.0 - 2.0 cm from the root tip. This change in cell morphology implied the formation of lysigenous aerenchyma in oil palm roots under waterlogging stress. At the same time, most of the candidate genes involved in lysigenous aerenchyma formation revealed a higher mRNA expression after waterlogging stress for 3 d. Genes of ethylene synthesis group ACS3, ACO, and ACO1 were highly up-regulated in both types of roots, while XTH22, XTH23, and CEL12 in the cell wall modification group were more highly up-regulated in the primary roots than in the secondary roots. CML11, CAMTA4, TCTP, and CPI1 in a signaling group were up-regulated in the primary roots, but they were down-regulated in the secondary roots. NAC29, ERF1, ERF113, and HSFA2C in a transcription factor group were strongly up-regulated in the oil palm roots. However, there have been no previous reports on the expression of CAMTA4, bHLH79, and bHLH94 under waterlogging conditions. Our findings confirm gene expression during lysigenous aerenchyma development in oil palm roots under waterlogging. It can also be stated that primary roots are an important part of the adaptation mechanism of oil palm roots for survival under waterlogging stress. Furthermore, the molecular markers of all expressed genes will be developed and applied in our oil palm breeding project for selection of waterlogging tolerance.
油棕(Elaeis guineensis Jacq.)通过诱导溶生通气组织来应对淹水胁迫,这有助于氧气通过其根组织流动,以在淹水条件下生存。因此,评价了油棕基因型Deli×Calabar在淹水胁迫下根系溶生通气组织形成的形态和遗传适应。本研究发现,淹水胁迫2 d后,次生根的死细胞数量最多,而根系孔隙率随时间的增加而增加,尤其是在距根尖1.0-2.0 cm处。这种细胞形态的变化暗示了在淹水胁迫下油棕根中溶生通气组织的形成。同时,大多数参与溶生通气组织形成的候选基因在淹水胁迫3天后显示出更高的mRNA表达。乙烯合成组ACS3、ACO和ACO1的基因在两种类型的根中都高度上调,而XTH22、XTH23、,细胞壁修饰组中的CEL12在初生根中比在次生根中更高上调。信号组中的CML11、CAMTA4、TCTP和CPI1在初级根中上调,但在次级根中下调。转录因子组中的NAC29、ERF1、ERF113和HSFA2C在油棕根中强烈上调。然而,以前没有关于CAMTA4、bHLH79和bHLH94在内涝条件下表达的报道。我们的研究结果证实了在淹水条件下油棕根溶生通气组织发育过程中的基因表达。也可以说,初生根是油棕根在淹水胁迫下生存适应机制的重要组成部分。此外,将开发所有表达基因的分子标记,并将其应用于我们的油棕育种项目,以选择耐涝性。
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引用次数: 2
Ectopic expression of GmNHX3 and GmNHX1, encoding two Glycine max Na+/H+ vacuolar antiporters, improves water deficit tolerance in Arabidopsis thaliana GmNHX3和GmNHX1的异位表达,编码两个甘氨酸最大Na+/H+液泡反转运蛋白,提高了拟南芥对缺水的耐受性
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-06-09 DOI: 10.32615/BP.2021.003
E. M. Pardo, L. Toum, L. S. Pérez-Borroto, L. Fleitas, J. P. Gallino, S. Machi, A. Vojnov, A. Castagnaro, B. Welin
The importance of Na+/H+ antiporters in salt tolerance in plants has been demonstrated in many studies, but much less is known about their protective role during drought stress. To study their possible contribution to water deficit tolerance, two closely related soybean Na+/H+ antiporters belonging to the intracellular NHX exchanger protein family, GmNHX3 and GmNHX1, were evaluated in transgenic Arabidopsis thaliana. A. thaliana plants ectopically expressing GmNHX3 or GmNHX1 displayed a more drought-tolerant phenotype compared to wild-type plants, which was accompanied by an increase in relative water content and chlorophyll content during stress conditions. Both GmHNX1 and GmHNX3 transgenic lines accumulated higher amounts of Na+ and K+ cations, showed increased antioxidant enzyme activities and less membrane damage due to lipid peroxidation under water deficit, as compared to non-transformed plants. Furthermore, plants expressing GmNHX3 showed an increased sensitivity to abscisic acid as deduced from stomatal closure and seed germination inhibition studies. Finally, a significant up-regulation of abiotic stress-related genes was observed in both transgenic lines compared to wild-type plants in response to abscisic acid and mannitol treatments. These results demonstrate that GmNHX3 and GmNHX1 antiporters confer protection during drought stress in A. thaliana and hence are potential genetic targets to improve drought tolerance in soybean and other crops.
Na+/H+反转运蛋白在植物耐盐性中的重要性已在许多研究中得到证实,但对其在干旱胁迫中的保护作用知之甚少。为了研究它们对水分亏缺耐受性的可能贡献,在转基因拟南芥中评估了属于细胞内NHX交换蛋白家族的两种密切相关的大豆Na+/H+反转运蛋白GmNHX3和GmNHX1。与野生型植物相比,外源表达GmNHX3或GmNHX1的拟南芥植物表现出更耐旱的表型,这伴随着在胁迫条件下相对含水量和叶绿素含量的增加。与未转化的植物相比,GmHNX1和GmHNX3转基因株系在缺水条件下积累了更高量的Na+和K+阳离子,表现出更强的抗氧化酶活性和更少的由于脂质过氧化引起的膜损伤。此外,从气孔关闭和种子发芽抑制研究推断,表达GmNHX3的植物对脱落酸的敏感性增加。最后,与野生型植物相比,在两个转基因系中观察到非生物胁迫相关基因对脱落酸和甘露醇处理的显著上调。这些结果表明,GmNHX3和GmNHX1反转运蛋白在拟南芥干旱胁迫期间提供保护,因此是提高大豆和其他作物抗旱性的潜在遗传靶标。
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引用次数: 0
Physiological and transcriptomic analysis of Pinus massoniana seedling response to osmotic stress 马尾松幼苗对渗透胁迫反应的生理和转录组学分析
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-05-31 DOI: 10.32615/BP.2021.016
H. Xu, X. Gao, C. Yu
Masson pine (Pinus massoniana Lamb.) is an important tree species of high economic value in southern China, but osmotic stress threatens its growth and development. In this study, physiological measurements and RNA-Seq analysis were used to clarify the physiological and molecular responses of P. massoniana under osmotic stress. Osmotic treatment caused cell membrane damage and reactive oxygen species (ROS) accumulation in the tree seedlings, but it also increased their antioxidant enzyme (superoxide dismutase, peroxidase, and catalase) activities and osmotic substances (soluble sugars, proline, and trehalose) content so as to adjust to osmotic stress conditions. A total of 1 789 differentially expressed genes (DEGs) were identified by transcriptome sequencing, of which 962 were up-regulated and 827 genes down-regulated. A series of stress-induced genes associated with signal transduction, ROS-scavenging, osmotic regulation, late embryogenesis abundant (LEA) protein, pentatricopeptide repeat-containing protein, and transcription factors' regulation were distinguishable. This detailed investigation of the stress-responsive genes and pathways provides new insight into molecular mechanism of abiotic stress response in P. massoniana. Further, this study's data can contribute to genetic engineering or molecular breeding efforts to enhance osmotic resistance in P. massoniana stands.
马尾松(Pinus massonian Lamb.)是我国南方重要的高经济价值树种,但渗透胁迫威胁着其生长发育。本研究采用生理学测量和RNA-Seq分析方法,阐明了马尾松在渗透胁迫下的生理和分子反应。渗透处理引起了细胞膜损伤和活性氧(ROS)在幼苗中的积累,但也增加了它们的抗氧化酶(超氧化物歧化酶、过氧化物酶和过氧化氢酶)活性和渗透物质(可溶性糖、脯氨酸和海藻糖)含量,以适应渗透胁迫条件。转录组测序共鉴定出1789个差异表达基因,其中962个上调,827个下调。可区分一系列与信号转导、ROS清除、渗透调节、晚期胚胎发生丰富蛋白(LEA)、含五肽重复序列蛋白和转录因子调节相关的应激诱导基因。对马尾松胁迫反应基因和途径的详细研究为揭示马尾松非生物胁迫反应的分子机制提供了新的见解。此外,本研究的数据可以为提高马尾松林分渗透阻力的基因工程或分子育种工作做出贡献。
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引用次数: 2
Comparison of antioxidant enzyme activity and gene expression in two new spring wheat cultivars treated with salinity 盐碱化处理下两个春小麦新品种抗氧化酶活性及基因表达的比较
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-05-17 DOI: 10.32615/BP.2020.171
Y. L. Xu, Yaolei Zhang, J. M. Li, Tianpeng Gao, L. N. Zhang, L. Si, Qiang Li, G. Li, Y. Yang
This study aimed to analyze element content, antioxidative response, and related gene expression in two new wheat (Tritium aestivum L.) cultivars Longchun 30 and Longchun 27 when exposed to different NaCl concentrations. Low NaCl concentration (25 mM) promoted root growth and decreased malondialdehyde (MDA) content and relative conductivity (REC) in Longchun 30. Differently, higher salinity stress (100 and 200 mM NaCl) inhibited root growth and increased MDA content and REC in both cultivars. Under salt stress, the increment of Na content in the roots and leaves and the reduction of Ca content in the roots were more remarkable in Longchun 27 than in Longchun 30. In contrast, the potassium content decreased in the roots but did not significantly change in the leaves in both cultivars under salinity. When the seedlings were exposed to salinity, the increases of superoxide dismutase (SOD) and catalase (CAT) activities in Longchun 27 roots were associated with high isoenzymes abundance and high TaCu/ZnSOD, TaMnSOD and TaCAT expression. Meanwhile, total peroxidase (POD) activity induced by NaCl treatment coincided with the changes of TaPOD expression and isoenzyme abundance in both cultivars. Besides, the inhibition of activities of apoplastic antioxidant enzymes, cell wall-bound POD, diamine oxidase, and polyamine oxidase was observed in salinity-stressed roots of both cultivars. Taken together, cv. Longchun 30 might be more suitable for growing in salinity environment in comparison with Longchun 27.
研究了小麦新品种龙春30号和龙春27号在不同NaCl浓度下的元素含量、抗氧化反应及相关基因表达。低NaCl浓度(25 mM)促进了龙春30根系生长,降低了丙二醛(MDA)含量和相对电导率(REC)。不同的是,高盐胁迫(100和200 mM NaCl)抑制了两个品种的根系生长,增加了MDA含量和REC。在盐胁迫下,龙春27的根和叶中Na含量的增加和根中Ca含量的降低比龙春30更为显著。盐胁迫下,两个品种的根钾含量下降,但叶片钾含量变化不显著。幼苗受盐胁迫后,龙春27根系超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性升高,同工酶丰度高,TaCu/ZnSOD、TaMnSOD和TaCAT表达量高。同时,NaCl处理诱导的总过氧化物酶(POD)活性与TaPOD表达量和同工酶丰度的变化一致。此外,两种品种的根外胞体抗氧化酶、细胞壁结合POD、二胺氧化酶和多胺氧化酶活性均受到抑制。综上所述,cv。与龙春27相比,龙春30可能更适合在盐碱环境下生长。
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引用次数: 2
Microtubule interaction of LICC1, a maize homologue of a component of the human muskelin/RanBPM/CTLH protein complex LICC1的微管相互作用,LICC1是人类滑鼠蛋白/RanBPM/CTLH蛋白复合物一种成分的玉米同源物
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-05-17 DOI: 10.32615/BP.2020.168
M. Miquel, D. Pagès-Vila, C. Vicient
MRCTLH (muskelin/RanBPM/CTLH) is a protein complex found in humans (MRCTLH) that is involved in the regulation of numerous cellular processes, such as gluconeogenesis, cell signaling, development, nuclear extrusion, cell morphology, or stability of different proteins. According to genomic data, all eukaryotes have similar protein complexes. In yeast, a similar protein complex named GID was found to be involved in the regulation of gluconeogenesis. LICC1 is a maize protein whose sequence resembles that of TWA1 in humans and GID8 in yeast, which are central components of the MRCTLH and GID complexes. LICC1 contains three highly conserved protein domains, LisH, CTLH, and CRA, typical of this protein family. Twa1 and gid8 are unique genes in human and yeast genomes. However, three copies of licc1 are present in the maize genome and multiple copies are present in other plant genomes. This result suggests the presence of multiple variants of the MRCTLH/GID complex in plants, which could increase its regulatory capacity. We also demonstrate here that LICC1 has the ability to interact with microtubules, similarly to the human TWA1. This interaction reinforces the idea that the LICC1 protein from maize, and its homologues in plants and, in general, the GID/MRCTLH complex in plants, can perform biological functions similar to those in humans and yeast.
MRCTLH (muskelin/RanBPM/CTLH)是一种在人类中发现的蛋白质复合物(MRCTLH),它参与许多细胞过程的调节,如糖异生、细胞信号传导、发育、核挤压、细胞形态或不同蛋白质的稳定性。根据基因组数据,所有真核生物都有相似的蛋白质复合物。在酵母中,一种名为GID的类似蛋白质复合物被发现参与糖异生的调节。LICC1是一种玉米蛋白,其序列类似于人类中的TWA1和酵母中的GID8,它们是MRCTLH和GID复合物的核心成分。LICC1包含三个高度保守的蛋白结构域,即LisH、CTLH和CRA,是该蛋白家族的典型结构域。Twa1和gid8是人类和酵母基因组中独特的基因。然而,licc1在玉米基因组中有3个拷贝,在其他植物基因组中有多个拷贝。这一结果表明植物中存在MRCTLH/GID复合体的多个变体,这可能会增加其调控能力。我们在这里也证明了LICC1具有与微管相互作用的能力,类似于人类的TWA1。这种相互作用强化了一种观点,即玉米中的LICC1蛋白及其在植物中的同源物,以及植物中的GID/MRCTLH复合物,可以发挥与人类和酵母相似的生物学功能。
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引用次数: 0
Tolerance of Arabidopsis thaliana plants overexpressing grapevine VaSTS1 or VaSTS7 genes to cold, heat, drought, salinity, and ultraviolet irradiation 过表达葡萄VaSTS1或VaSTS7基因的拟南芥植物对冷、热、干旱、盐度和紫外线照射的耐受性
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-05-17 DOI: 10.32615/BP.2020.139
Z. Ogneva, O. A. Aleynova, A. R. Suprun, Y. Karetin, A. S. Dubrovina, K. Kiselev
Stilbene synthases (STS) are plant enzymes that are responsible for the biosynthesis of stilbenes, which are plant phenolic compounds with valuable biological properties. Stilbenes also play important roles in plant tolerance to biotic and abiotic stresses. Therefore, plants that overexpress STS genes can be more resistant to various stresses. This paper investigated the effects of STS gene overexpression in Arabidopsis thaliana (L.) Heynh. Columbia-0 plants on stilbene content and tolerance to the following abiotic stresses: low and high temperatures, salinity, drought, and ultraviolet irradiation (UV-B and UV-C). We used VaSTS1 and VaSTS7 genes from grapevine (Vitis amurensis Rupr.) expressed under the double cauliflower mosaic virus 35S (CaMV35S) promoter. This study firstly demonstrated that overexpression of the VaSTS1 and VaSTS7 genes in A. thaliana plants considerably increased plant tolerance to UV-B and UV-C, while the tolerance to the low and high temperatures, salinity, and drought was not affected. We showed that the highest trans-piceid and trans-resveratrol total content was in ST1 A. thaliana plants that overexpressed the VaSTS1 gene in the range 8.28 - 22.66 µg g-1(f.m.). ST7 plants that overexpressed the VaSTS7 gene showed only trans-resveratrol at 0.02 - 0.08 µg g-1(f.m). Stilbene content and UV tolerance in transgenic A. thaliana plants correlated with STS transgene expression. STS expression, UV tolerance, and stilbene content was higher in VaSTS1 transgenic plants compared with that in VaSTS7 transgenic plants.
二苯乙烯合成酶(STS)是负责二苯乙烯生物合成的植物酶,二苯乙烯是一种具有宝贵生物学特性的植物酚类化合物。二苯乙烯在植物对生物和非生物胁迫的耐受性中也发挥着重要作用。因此,过表达STS基因的植物可以对各种胁迫更有抵抗力。本文研究了STS基因在拟南芥(Arabidopsis thaliana(L.)Heynh中过表达的影响。哥伦比亚-0植物对二苯乙烯含量和对以下非生物胁迫的耐受性:低温和高温、盐度、干旱和紫外线照射(UV-B和UV-C)。我们使用了在双花椰菜花叶病毒35S(CaMV35S)启动子下表达的葡萄(Vitis amurensis Rupr.)的VaSTS1和VaSTS7基因。本研究首次证明,VaSTS1和VaSTS7基因在拟南芥中的过表达显著提高了植物对UV-B和UV-C的耐受性,而对低温和高温、盐度和干旱的耐受性不受影响。我们发现,反式苦味酸和反式白藜芦醇总含量最高的是过表达VaSTS1基因的ST1拟南芥植物,其范围为8.28-22.66µg g-1(f.m。与VaSTS7转基因植物相比,VaSTS1转基因植物的STS表达、紫外线耐受性和二苯乙烯含量更高。
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引用次数: 1
Rapid and efficient leaf regeneration propagation system for Euonymus bungeanus 白卫矛叶片再生快速高效繁殖系统
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-05-17 DOI: 10.32615/BP.2020.166
K. Ning, T. Zhou, C. Jiang, WU H.M., J. Jiang, J. Chen, Y. El-Kassaby, M. Y
Rapid propagation of plants by tissue culture is of great significance for large-scale production, molecular genetics research, and breeding. Currently, a rapid and high-efficient tissue culture protocol for Euonymus bungeanus is needed. To develop a propagation system for this species, we established a new regeneration system from leaves. Callus formation was induced on Murashige and Skoog (MS) medium supplemented with 0.5 mg dm-3 6-benzylaminopurine (6-BA) and 0.5 mg dm-3 α-naphthalene acetic acid (NAA) and the induction rate almost reached 100 % under red radiation within 21 d. The medium for proliferation of adventitious buds comprised of MS medium with 1.0 mg dm-3 6-BA and 0.5 mg dm-3 NAA, and the induction rate within 20 d nearly reached 100 %. When, the adventitious buds were transferred to the rooting medium containing 1/2MS, 2.0 mg dm-3 indole-3-butyric acid (IBA), and 0.05 mg dm-3 NAA, adventitious root formation was achieved within 20 d. Collectively, the rapid and high-efficient regeneration system from E. bungeanus leaves was established, providing useful references for effective mass propagation and it could serve as an enabling technology for future genetic engineering.
植物组织培养快速繁殖对大规模生产、分子遗传学研究和育种具有重要意义。目前,需要一种快速高效的白芷组织培养方案。为了开发该物种的繁殖系统,我们建立了一个新的叶片再生系统。在添加0.5mg dm-3 6-苄基氨基嘌呤(6-BA)和0.5mg dm3α-萘乙酸(NAA)的Murashige和Skoog(MS)培养基上诱导愈伤组织形成,在红色辐射下21 d内诱导率几乎达到100%,20d内诱导率接近100%。当不定芽转移到含有1/2MS、2.0mg dm-3吲哚-3-丁酸(IBA)和0.05mg dm-3 NAA的生根培养基中时,不定根在20d内形成,为有效的大规模繁殖提供了有益的参考,可以作为未来基因工程的一种使能技术。
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引用次数: 1
Influence of water shortage on apple seedling growth under different radiation composition 不同辐射成分下缺水对苹果幼苗生长的影响
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-05-12 DOI: 10.32615/BP.2020.086
F. A. Hamann, A. Fiebig, G. Noga
Abiotic stresses strongly impair plant development and might impose detrimental effects particularly on seedlings. Irradiance and water deficit are relevant factors, which affect performance of young plants under controlled conditions. In our study, we investigated the influence of water shortage combined with different radiation sources - light emitting diodes (LED) and compact fluorescence lamps (CFL) - on physiological and biochemical parameters of young apple plants. Stress responses were assessed by fluorescence-based indices, while relative water, chlorophyll (Chl), and proline content served as reference parameters. The watering regime had a higher impact on biochemical indicators than the radiation sources. Lower Chl content was determined in plants grown under LED both in control and in water deficit plants. Nitrogen balance index and nitrogen balance index with red radiation excitation showed similar patterns regarding leaf Chl results in relation to the radiation source, being higher under CFL. In contrast, the flavonol index was higher in plants cultivated under LED. Stomatal conductance and maximal photochemical efficiency emphasised a radiation quality effect with higher values for CFL. In conclusion, fluorescence indices related to nitrogen status and flavonol content are promising parameters to sense physiological impairments under the given conditions. However, discrepancies compared to previous studies might be related to the different plant species, the nature of dehydration, and the measuring conditions.
非生物胁迫严重损害植物发育,并可能对幼苗产生不利影响。光照和水分亏缺是影响苗木生长发育的重要因素。本研究研究了不同光源(LED和CFL)在缺水条件下对苹果幼苗生理生化参数的影响。以相对水分、叶绿素(Chl)和脯氨酸含量为参考参数,采用荧光指标评价胁迫反应。灌水制度对生化指标的影响大于辐射源。在LED下生长的对照植株和水分亏缺植株的Chl含量均较低。与辐射源相关的叶片Chl结果中,氮平衡指数与红色辐射激发下的氮平衡指数表现出相似的规律,在CFL下更高。相比之下,在LED下栽培的植株黄酮醇指数更高。气孔导度和最大光化学效率强调了CFL较高的辐射质量效应。综上所述,在一定条件下,与氮状态和黄酮醇含量相关的荧光指标是有希望检测生理损伤的参数。然而,与以往研究相比,差异可能与不同的植物种类、脱水性质和测量条件有关。
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引用次数: 0
Topical application of double-stranded RNA molecules deriving from Tomato yellow leaf curl virus reduces cognate virus infection in tomato 番茄黄叶卷曲病毒双链RNA分子的局部应用减少番茄同源病毒感染
IF 1.5 4区 生物学 Q2 Agricultural and Biological Sciences Pub Date : 2021-05-12 DOI: 10.32615/BP.2020.172
O. Melita, A. Kaldis, M. Berbati, C. Reppa, M. Holeva, M. Lapidot, D. Gelbart, P. Otten, A. Voloudakis
Tomato yellow leaf curl virus (TYLCV), a whitefly-transmitted single-stranded DNA (ssDNA) virus, causes the most important viral disease of tomato worldwide. TYLCV-mediated disease is mainly controlled via extensive insecticide sprays aiming at the whitefly vector. RNA-based vaccination was proven to be a non-transgenic approach leading to efficient plant virus control. In this work, double-stranded RNA (dsRNA) molecules deriving from sequences of the C4 and V2 genes of TYLCV-Mild were produced in vitro and topically applied onto tomato plants along with the virus (via agroinfiltration). DsC4 and dsV2 application reduced disease incidence to 23 and 46 %, respectively, while TYLCV positive control reached 64 %. Bioinformatics analysis of the virus-specific small interfering RNAs (vsiRNAs) from TYLCV-infected tomato revealed 'hot' and 'cold' spots in the TYLCV-Mild genome. Interestingly, the viral C-strand had twofold siRNA reads when compared to that of the V-strand. Overall, vsiRNAs of negative and positive polarity were almost equal (53.5 vs. 46.6 %); vsiRNAs of negative polarity prevailed at the V-strand. Stem-loop RT-PCR validated the presence of six vsiRNAs (hot or cold spots) in TYLCV-Mild-infected and dsRNA-treated tomato. The exogenously applied dsRNA was found to rapidly move systemically in tomato and was detected for 54 days post treatment (dpt). The applied dsRNA molecules were successfully processed by the Dicer-like proteins (DCLs) in tomato since small interfering RNAs (siRNAs) deriving from the dsRNA were detected for at least 54 dpt. This consists the first report of dsRNA-based vaccination applied against a monopartite geminivirus.
番茄黄叶卷曲病毒(TYLCV)是一种由白蝇传播的单链DNA(ssDNA)病毒,是世界范围内番茄最重要的病毒性疾病。TYLCV介导的疾病主要通过针对粉虱媒介的广泛杀虫剂喷雾来控制。基于RNA的疫苗接种已被证明是一种非转基因方法,可有效控制植物病毒。在这项工作中,来源于TYLCV温和型C4和V2基因序列的双链RNA(dsRNA)分子在体外产生,并与病毒一起(通过农业渗透)局部应用于番茄植株上。DsC4和dsV2的应用将疾病发生率分别降低到23%和46%,而TYLCV阳性对照达到64%。对TYLCV感染番茄的病毒特异性小干扰RNA(vsiRNA)的生物信息学分析揭示了TYLCV轻度基因组中的“热点”和“冷点”。有趣的是,与V链相比,病毒C链的siRNA读数是V链的两倍。总体而言,负极性和正极性的vsiRNA几乎相等(53.5%对46.6%);负极性的vsiRNA在V链上占主导地位。茎环RT-PCR验证了TYLCV轻度感染和dsRNA处理的番茄中存在六个vsiRNA(热点或冷点)。发现外源应用的dsRNA在番茄中快速系统移动,并在处理后54天内检测到(dpt)。番茄中的Dicer样蛋白(DCLs)成功地处理了所应用的dsRNA分子,因为从dsRNA衍生的小干扰RNA(siRNA)被检测到至少54dpt。这是第一份基于dsRNA的疫苗接种单体双子病毒的报告。
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引用次数: 9
期刊
Biologia Plantarum
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