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Arsenic tolerance unveiled in Arabidopsis: CPK23 and PHT1;1 alliance 揭示拟南芥的砷耐受性:CPK23 和 PHT1;1 联盟
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-21 DOI: 10.1007/s13562-024-00885-1
Jawahar Singh, Vishnu Mishra, Vishal Varshney

Arsenate As(V), characterized as a metalloid with heavy metal properties, is prevalent in various environments. The consumption of food derived from plants contaminated with arsenate contributes significantly to human exposure to arsenic, posing potential health risks. However, the mechanisms governing plant responses to arsenate stress and the regulation of relevant transporter functions remain inadequately understood. Recently, Wang and co-workers, identified a calcium-dependent protein kinase, specifically CALCIUM-DEPENDENT PROTEIN KINASE 23 (CPK23), which shows interaction with the plasma membrane As(V)/Pi transporter PHOSPHATE TRANSPORTER 1;1 (PHT1;1) to channelize the Ca2+ signal in Arabidopsis roots under As(V) stress. The authors observed that cpk23 mutants showed increased sensitivity, whereas the overexpression of CPK23 resulted in enhanced tolerance under As(V) stress conferring role in As stress. Moreover, it has been demonstrated that CPK23 phosphorylates PHT1.1 at the Ser514 (S514) site is crucial for its function and proper localization under As(V) stress. Thus, this commentary offers valuable insights into the induction of a notable Ca2+ signal in Arabidopsis roots under As(V) stress that could guide crop bioengineering efforts aimed at addressing arsenate pollution in soil with targeted strategies.

砷酸盐 As(V) 是一种具有重金属特性的类金属,普遍存在于各种环境中。食用来自受砷酸盐污染的植物的食物大大增加了人类接触砷的机会,对健康构成潜在风险。然而,人们对植物对砷酸盐胁迫的反应机制以及相关转运体功能的调控仍缺乏足够的了解。最近,Wang 和合作者发现了一种钙依赖性蛋白激酶,特别是钙独立蛋白激酶 23(CPK23),它与质膜 As(V)/Pi 转运体 PHOSPHATE TRANSPORTER 1;1 (PHT1;1) 相互作用,在拟南芥根中引导 As(V) 胁迫下的 Ca2+ 信号。作者观察到,cpk23 突变体对砷胁迫的敏感性增加,而过表达 CPK23 则增强了对砷胁迫的耐受性。此外,研究还证明 CPK23 在 Ser514(S514)位点磷酸化 PHT1.1,这对其在 As(V)胁迫下的功能和正确定位至关重要。因此,这篇评论对拟南芥根系在As(V)胁迫下诱导显著的Ca2+信号提供了有价值的见解,可以指导作物生物工程工作,以有针对性的策略解决土壤中的砷酸盐污染问题。
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引用次数: 0
Identification of superior haplotypes for seed protein content in pigeonpea (Cajanus cajan L.) 鉴定鸽子豆(Cajanus cajan L.)种子蛋白质含量的优良单倍型
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-18 DOI: 10.1007/s13562-024-00884-2
Harsha Vardhan Rayudu Jamedar, Prasad Gandham, Prasad Bajaj, Srinivas Thati, V. Srinivasa Rao, Rajeev K. Varshney, Rachit K. Saxena

Pigeonpea (Cajanus cajan L.) is an important source of quality dietary protein for over a billion people worldwide. The seeds of pigeonpea contain approximately 20–22% digestible protein, which makes it a valuable source of nutrition. Despite this, there has been little attention paid to enhancing the seed protein content (SPC) through genetic means. Recently, high-protein germplasm lines have been discovered in the secondary gene pool, which presents an opportunity to breed for high-protein cultivars. To accelerate the breeding process, genomics-assisted breeding (GAB) can be utilized. In this context, this study identified the superior haplotypes for the genes that control SPC in pigeonpea. Whole-genome re-sequencing (WGRS) data from 344 pigeonpea genotypes were analyzed to identify the superior haplotypes for 57 SPC governing genes. A total of 231 haplotypes in 43 candidate genes were identified, and haplo-pheno analysis was performed to provide superior haplotypes for 10 genes. The identification of superior haplotypes and genotypes will greatly facilitate the development of protein-rich pigeonpea seeds through the application of haplotype-based breeding (HBB).

鸽子豆(Cajanus cajan L.)是全球十多亿人优质膳食蛋白质的重要来源。鸽子豆的种子含有约 20-22% 的可消化蛋白质,这使其成为一种宝贵的营养来源。尽管如此,人们很少关注通过遗传手段提高种子蛋白质含量(SPC)。最近,在次级基因库中发现了高蛋白种质系,这为培育高蛋白栽培品种提供了机会。为了加快育种进程,可以利用基因组辅助育种(GAB)。在此背景下,本研究确定了控制鸽子豆 SPC 基因的优势单倍型。通过分析 344 个鸽子豆基因型的全基因组重测序(WGRS)数据,确定了 57 个 SPC 调控基因的优良单倍型。共鉴定出 43 个候选基因中的 231 个单倍型,并对 10 个基因进行了单倍型-表型分析,以提供优良的单倍型。优良单倍型和基因型的鉴定将极大地促进通过应用基于单倍型的育种(HBB)来培育富含蛋白质的鸽子豆种子。
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引用次数: 0
Enhanced drought and salt tolerance of Arabidopsis thaliana by ectopic expression of the molecular chaperone artemin from Artemia urmiana 通过异位表达来自蒿属植物的分子伴侣蛋白青蒿素增强拟南芥的耐旱和耐盐能力
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-16 DOI: 10.1007/s13562-024-00877-1
Zeinab Poormohammad, Sara Shahrokhi, Amin Abedi, Reza H. Sajedi, M. Mehdi Sohani

Due to the resistance of Artemia urmiana to salt stress, researchers have isolated and investigated Artemin, the most prevalent protein within the cyst of this aquatic species. In vitro studies have revealed Artemin's role as a molecular chaperone, effectively engaging with the hydrophobic surfaces of unfolded and/or partially folded proteins. In light of Artemin's established functional significance, its encoding gene has been successfully introduced into mammalian cells; however, no published research has elucidated its potential role within plant cells. In the current investigation, the artemin gene was successfully cloned into the pPZPY122 plant vector and subsequently introduced into Arabidopsis thaliana plants. The T3 homozygote transgenic plants (art) were then subjected to a series of environmental stresses, including heat, salt (NaCl) and drought (Mannitol). To assess the mutant's resilience to these stresses, their seed germination indices were evaluated. The art line demonstrated a higher degree of tolerance towards the abiotic stresses. A comparative analysis revealed that ascorbate peroxidase activity, catalase activity, and proline content exhibited significantly enhanced levels in some NaCl-treated art plants compared to their counterparts in Col-0. Regarding the expression of the genes in the SOS pathway, it was found that SOS1 is significantly upregulated under NaCl treatment in the art mutant. Conversely, under normal growth conditions, the morphology and growth of transgenics remained indistinguishable from those of wild-type plants.

由于 Artemia urmiana 对盐胁迫具有抗性,研究人员分离并研究了 Artemin,这是这种水生物种包囊中最常见的蛋白质。体外研究揭示了 Artemin 的分子伴侣作用,它能有效地与未折叠和/或部分折叠蛋白质的疏水表面接触。鉴于青蒿素的既定功能意义,其编码基因已被成功导入哺乳动物细胞;然而,尚未有公开发表的研究阐明其在植物细胞中的潜在作用。在本次研究中,青蒿素基因被成功克隆到 pPZPY122 植物载体中,随后被导入到拟南芥植株中。然后将 T3 同源基因转基因植株(art)置于一系列环境胁迫下,包括热、盐(氯化钠)和干旱(甘露醇)。为了评估突变体对这些胁迫的适应能力,对它们的种子萌发指数进行了评估。艺术系对非生物胁迫表现出更高的耐受性。比较分析表明,与 Col-0 的同类基因相比,经过 NaCl 处理的艺术系植株的抗坏血酸过氧化物酶活性、过氧化氢酶活性和脯氨酸含量都明显提高。在 SOS 通路基因的表达方面,研究发现 SOS1 在 NaCl 处理下明显上调。相反,在正常生长条件下,转基因植株的形态和生长与野生型植株没有区别。
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引用次数: 0
Genetic diversity assessment of groundnut (Arachis hypogaea L.) for polyphenol content and antioxidant activity: unlocking the nutritional potential 花生(Arachis hypogaea L.)多酚含量和抗氧化活性的遗传多样性评估:挖掘营养潜力
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-15 DOI: 10.1007/s13562-024-00882-4
Spoorti S. Gandhadmath, S. Vidyashree, Rakeshkumar Choudhary, Babu N. Motagi, Ravikumar Hosamani, Pushpa Bharati, Spurthi N. Nayak

Groundnut is a rich source of several nutritional components including polyphenols and antioxidants that offer various health benefits. In this regard, the mini core accessions along with elite varieties of groundnut were used to assess genetic diversity using AhTE markers. The phenotypic observation on eight morphological, six productivity and two nutraceutical traits [total polyphenol content (TPC) and total antioxidant activity (AOA)] were studied. Correlation studies revealed a significant positive correlation between TPC and AOA. The degree of divergence with respect to nutraceutical content among the genotypes of mini core collection and elite cultivars is evident from the current study. The STRUCTURE analyses revealed the grouping of genotypes into three distinct clusters mainly based on the botanical types of groundnut. The analysis of molecular variance displayed maximum variation (97%) within the individuals of subpopulations and minimum variation (3%) among subpopulations. Principal component analysis exhibited 3 principal components that accounted for 42.17% of the total variation. Association mapping study indicated 20 significant marker-trait associations at 1% probability. The study has also identified significant marker-trait associations with nutraceutical properties of groundnut, AhTE0465-TPC and AhTE0381- AOA with explained phenotypic variation of 7.45% and 6.85% respectively. These markers were found to have positions at A02 and A09 with bHLH DNA-binding family protein and chitinase putative functions respectively. The markers associated with TPC and AOA can further be utilized for genomics-assisted breeding for nutritionally rich cultivars in groundnut.

落花生富含多种营养成分,包括多酚和抗氧化剂,对健康有多种益处。为此,利用 AhTE 标记对小型核心登录品种和花生优良品种的遗传多样性进行了评估。研究了八个形态特征、六个产量特征和两个营养保健性状(总多酚含量(TPC)和总抗氧化活性(AOA))的表型观察。相关研究表明,总多酚含量和总抗氧化活性之间存在明显的正相关。从目前的研究中可以明显看出,迷你核心品系和精英栽培品种的基因型在营养保健成分方面的差异程度。结构分析表明,基因型主要根据落花生的植物学类型分为三个不同的群组。分子变异分析表明,亚群个体内部的变异最大(97%),亚群之间的变异最小(3%)。主成分分析显示有 3 个主成分,占总变异的 42.17%。关联图谱研究表明,在 1%的概率下,有 20 个显著的标记-性状关联。研究还发现,AhTE0465-TPC 和 AhTE0381- AOA 与落花生的营养保健特性有重要的标记-性状关联,解释的表型变异分别为 7.45% 和 6.85%。这些标记的A02和A09位置分别具有bHLH DNA结合家族蛋白和几丁质酶的推定功能。与 TPC 和 AOA 相关的标记可进一步用于花生营养丰富栽培品种的基因组辅助育种。
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引用次数: 0
Electrical Status Epilepticus during Sleep in a Male Filipino with Rare Nonsense Mutation Variant of Sotos Syndrome on Carbamazepine Monotherapy. 一名患有罕见无义突变变异型索托斯综合征的菲律宾男性在接受卡马西平单药治疗后出现睡眠时电癫痫状态。
4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-15 eCollection Date: 2024-01-01 DOI: 10.47895/amp.vi0.3013
Jeffrey I Lappay, Benilda C Sanchez-Gan, Michelle E Abadingo

Electrical status epilepticus during sleep (ESES) is an electrographic pattern associated with specific genetic disorders, brain malformations, and use of some antiseizure medications. This case report aims to present the management of ESES in Sotos syndrome (SoS) on carbamazepine. A nine-year-old Filipino male with clinical features suggestive of overgrowth syndrome presented with febrile seizure at one year old. Cranial imaging showed cavum septum pellucidum, corpus callosal dysgenesis, and ventriculomegaly. He was on carbamazepine monotherapy starting at three years old. A near continuous diffuse spike-wave discharges in slow wave sleep was recorded at nine years old hence shifted to valproic acid. Follow-up study showed focal epileptiform discharges during sleep with disappearance of ESES. Next generation sequencing tested positive for rare nonsense mutation of nuclear receptor binding set-domain protein 1 confirming the diagnosis of SoS. Advanced molecular genetics contributed to determination of ESES etiologies. To date, this is the first documented case of SoS developing ESES. Whether an inherent genetic predisposition or drug-induced, we recommend the avoidance of carbamazepine and use of valproic acid as first-line therapy.

睡眠中电癫痫状态(ESES)是一种与特定遗传疾病、脑部畸形和服用某些抗癫痫药物有关的电图模式。本病例报告旨在介绍服用卡马西平的索托斯综合征(SoS)患者的 ESES 治疗情况。一名九岁的菲律宾男性患者在一岁时出现发热性癫痫发作,临床特征提示为生长过度综合征。头颅影像学检查显示有透明隔腔、胼胝体发育不良和脑室肥大。他从三岁开始接受卡马西平单药治疗。九岁时在慢波睡眠中记录到近乎连续的弥漫性棘波放电,因此转为丙戊酸治疗。随访研究显示,睡眠中出现局灶性癫痫样放电,ESES消失。下一代测序结果显示,核受体结合集域蛋白1的罕见无义突变呈阳性,确诊为SoS。先进的分子遗传学有助于确定 ESES 的病因。迄今为止,这是有据可查的第一例SoS发展为ESES的病例。无论是固有的遗传倾向还是药物诱导,我们都建议避免使用卡马西平,并将丙戊酸作为一线治疗药物。
{"title":"Electrical Status Epilepticus during Sleep in a Male Filipino with Rare Nonsense Mutation Variant of Sotos Syndrome on Carbamazepine Monotherapy.","authors":"Jeffrey I Lappay, Benilda C Sanchez-Gan, Michelle E Abadingo","doi":"10.47895/amp.vi0.3013","DOIUrl":"10.47895/amp.vi0.3013","url":null,"abstract":"<p><p>Electrical status epilepticus during sleep (ESES) is an electrographic pattern associated with specific genetic disorders, brain malformations, and use of some antiseizure medications. This case report aims to present the management of ESES in Sotos syndrome (SoS) on carbamazepine. A nine-year-old Filipino male with clinical features suggestive of overgrowth syndrome presented with febrile seizure at one year old. Cranial imaging showed cavum septum pellucidum, corpus callosal dysgenesis, and ventriculomegaly. He was on carbamazepine monotherapy starting at three years old. A near continuous diffuse spike-wave discharges in slow wave sleep was recorded at nine years old hence shifted to valproic acid. Follow-up study showed focal epileptiform discharges during sleep with disappearance of ESES. Next generation sequencing tested positive for rare nonsense mutation of nuclear receptor binding set-domain protein 1 confirming the diagnosis of SoS. Advanced molecular genetics contributed to determination of ESES etiologies. To date, this is the first documented case of SoS developing ESES. Whether an inherent genetic predisposition or drug-induced, we recommend the avoidance of carbamazepine and use of valproic acid as first-line therapy.</p>","PeriodicalId":16835,"journal":{"name":"Journal of Plant Biochemistry and Biotechnology","volume":"29 1","pages":"83-87"},"PeriodicalIF":0.0,"publicationDate":"2024-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11219534/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70452246","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Inadequate lysine content of wheat endosperm proteins - possibility of correcting it by CRISPR-Cas system of genome editing 小麦胚乳蛋白质中赖氨酸含量不足--通过 CRISPR-Cas 基因组编辑系统纠正的可能性
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-12 DOI: 10.1007/s13562-024-00881-5
Hirendra Kumar Das, Megha Kaushik, Pranab Kumar Mandal

Proteins in endosperms of mature wheat seeds contain ~ 2.5% lysine. Proteins in endosperms of immature wheat seeds after 8 days of anthesis, interestingly contain ~ 8% lysine. Wheat endosperm proteins collected 15 days after anthesis, when treated with proteases isolated from endosperms 20 days after anthesis, released 10.2% lysine and 5.6% leucine. In contrast, the same proteins, when treated with proteases isolated from endosperms 8 days after anthesis, released 7.0% lysine and 12.0% leucine. The protease C, that could cleave polylysine, but could not cleave polyaspartic acid and polyglutamic acid, appeared only at 16 days after anthesis and continued to be present till 28 days after anthesis. Soybean trypsin inhibitor inhibited the activity of protease C completely. Transcriptome analysis of developing wheat seeds has revealed the presence of three major transcripts containing trypsin-like domains. The amounts of these three transcripts increased from 2 weeks after anthesis to 4 weeks after anthesis and decreased thereafter. If a wheat variety could be created which is devoid of protease C or the above mentioned trypsin like proteases, the lysine content of endosperm proteins of that variety would possibly be higher. This could today be achieved by using the CRISPR-Cas systems of genome editing.

成熟小麦种子胚乳中的蛋白质含有 ~ 2.5% 的赖氨酸。有趣的是,开花 8 天后未成熟小麦种子胚乳中的蛋白质含有 ~ 8%的赖氨酸。用从开花后 20 天的胚乳中分离出来的蛋白酶处理开花后 15 天收集的小麦胚乳蛋白质时,可释放出 10.2% 的赖氨酸和 5.6% 的亮氨酸。相反,用从花后 8 天的胚乳中分离出来的蛋白酶处理同样的蛋白质时,会释放出 7.0% 的赖氨酸和 12.0% 的亮氨酸。蛋白酶 C 能裂解聚赖氨酸,但不能裂解聚天冬氨酸和聚谷氨酸,它在花后 16 天才出现,并一直持续到花后 28 天。大豆胰蛋白酶抑制剂完全抑制了蛋白酶 C 的活性。对发育中的小麦种子进行的转录组分析表明,存在三种含有胰蛋白酶样结构域的主要转录本。这三种转录本的数量从开花后 2 周增加到 4 周,之后又有所减少。如果能培育出一种不含蛋白酶 C 或上述类似胰蛋白酶的小麦品种,该品种胚乳蛋白中的赖氨酸含量可能会更高。如今,利用 CRISPR-Cas 基因组编辑系统就可以实现这一目标。
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引用次数: 0
Molecular differentiation of the green and purple Tulsi (Ocimum tenuiflorum L.) and its application in authentication of market samples 绿色和紫色 Tulsi(Ocimum tenuiflorum L.)的分子分化及其在市场样本鉴定中的应用
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-11 DOI: 10.1007/s13562-024-00883-3
Mohamed Shaibi, Raju Balaji, Madasamy Parani

Tulsi (Holi Basil, Ocimum tenuiflorum) is extensively used in herbal medicine, and it includes two distinct subtypes; namely green Tulsi and purple Tulsi. Both types have similar medicinal properties. However, purple Tulsi contains a significantly higher amount of methyleugenol, which is genotoxic, and its daily intake is restricted. We developed a polymerase chain reaction-restriction fragment length polymorphism (PCR–RFLP) method to identify purple Tulsi. For this purpose, we selected a C > T single nucleotide polymorphism (SNP) in the ycf1 gene of the chloroplast genome that overlapped with a DdeI restriction site. The primers and PCR conditions were optimized to amplify a 797 bp DNA encompassing the C > T SNP, specifically from O. tenuiflorum. After restriction digestion of the PCR product with DdeI, green Tulsi was identified by two fragments (539 bp and 258 bp), and purple Tulsi was recognized by a single fragment (797 bp). Analysis of 40 Tulsi market samples revealed that only 36 (90%) were derived from O. tenuiflorum. The majority of the market samples were purple Tulsi (60%) or a mixture of green and purple Tulsi (27.5%), with some mixed samples containing up to 50% purple Tulsi.

图尔西(霍利罗勒,Ocimum tenuiflorum)被广泛用于草药,它包括两种不同的亚型,即绿色图尔西和紫色图尔西。这两种植物的药效相似。然而,紫图尔西含有较高的甲基丁香酚,具有遗传毒性,因此限制了其每日摄入量。我们开发了一种聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)方法来鉴定紫色图尔西。为此,我们选择了叶绿体基因组 ycf1 基因中与 DdeI 限制位点重叠的 C > T 单核苷酸多态性(SNP)。对引物和 PCR 条件进行了优化,以扩增包含 C > T SNP 的 797 bp DNA,特别是来自 O. tenuiflorum 的 DNA。用 DdeI 对 PCR 产物进行限制性消化后,通过两个片段(539 bp 和 258 bp)识别出绿色图尔西,通过一个片段(797 bp)识别出紫色图尔西。对 40 个图尔西市场样品的分析表明,只有 36 个(90%)来自 O. tenuiflorum。大多数市场样品是紫色图尔西(60%)或绿色和紫色图尔西的混合物(27.5%),一些混合样品中紫色图尔西的含量高达 50%。
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引用次数: 0
CRISPR/Cas system for the traits enhancement in potato (Solanum tuberosum L.): present status and future prospectives 用于马铃薯(Solanum tuberosum L.)性状改良的 CRISPR/Cas 系统:现状与前景
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-05 DOI: 10.1007/s13562-024-00878-0

Abstract

The increasing global population projected to reach 9.7 billion by 2050 from the current 7.7 billion, which is going to significantly impact food availability. Therefore, current global agricultural production needs to be increased to feed the unconstrained growing population. The changing climatic condition due to anthropogenic activities also makes the conditions more challenging to meet the required crop productivity in the future. Potato is third most consumed staple food. The current cultivation and demand of potato is particularly more in developing countries with high levels of poverty, hunger, and malnutrition because it is cheap source of nutrition and easily available. Potato is severely infected by a number of diseases, insect pests, and abiotic environmental conditions. Amidst the backdrop of climate change, the situation is deteriorating. Projections indicate that the average potato productivity in India's primary potato-growing states, responsible for approximately 90% of the nation's total potato output, is expected to decline by 2.0% in the 2050s and a more substantial 6.4% in the 2080s. Therefore to develop tolerance in plants to cope with changing environmental condition is need of hour to feed the overgrowing population. In this review, we discussed the application of CRISPR to enhance the crop productivity and develop biotic and abiotic stress-tolerant in potato to face the current changing climatic conditions is also discussed. The review also sheds light on the future prospects of CRISPR/Cas technology for potato breeding, including potential challenges and opportunities.

摘要 预计到 2050 年,全球人口将从目前的 77 亿增加到 97 亿,这将对粮食供应产生重大影响。因此,需要提高当前的全球农业产量,以养活不断增长的人口。人为活动导致的气候条件变化也使未来满足作物生产要求的条件更具挑战性。马铃薯是消费量第三大的主食。目前,马铃薯的种植和需求在贫困、饥饿和营养不良程度较高的发展中国家尤为突出,因为马铃薯是廉价的营养来源,而且很容易获得。马铃薯受到多种疾病、虫害和非生物环境条件的严重影响。在气候变化的背景下,情况正在恶化。预测表明,印度主要马铃薯种植邦的平均马铃薯产量(约占全国马铃薯总产量的90%)预计将在2050年代下降2.0%,到2080年代将大幅下降6.4%。因此,当务之急是开发植物的耐受性,以应对不断变化的环境条件,从而养活日益增多的人口。在这篇综述中,我们讨论了如何应用CRISPR来提高作物产量,并开发耐生物和非生物胁迫的马铃薯,以应对当前不断变化的气候条件。综述还揭示了CRISPR/Cas技术在马铃薯育种方面的未来前景,包括潜在的挑战和机遇。
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引用次数: 0
Measurement of transgene copy number in transgenic tobacco plants producing human interferon-γ (hIFN-γ) using quantitative real-time PCR 利用实时定量 PCR 测量产生人干扰素-γ(hIFN-γ)的转基因烟草植物的转基因拷贝数
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-04 DOI: 10.1007/s13562-024-00879-z
Reza Heidari Japelaghi, Raheem Haddad, Mostafa Valizadeh, Ebrahim Dorani Uliaie, Mokhtar Jalali Javaran

In transgenic plants, the transgene copy numbers can highly affect the level of expression and genetic stability of the transgene. Hence, the first step in their characterization is the estimation of transgene copy numbers integrated in the plant genome. Quantitative real-time PCR (qRT-PCR) was used to determine the copy numbers of human interferon-γ (hIFN-γ) and hygromycin phosphortransferase II (hptII) transgenes in the genome of the T0 generation of 18 transgenic tobacco lines using the axi1 gene as an endogenous control. With optimized PCR conditions, we attained highly exact estimates of one, two, three, and/or four transgene copies in the T0 transformants. Moreover, estimation of copy numbers of the hIFN-γ transgene and the hptII selective marker gene indicated that rearrangements of the T-DNA has regularly happened in transgenic tobacco. Transgene copy number was also estimated using Southern blot analysis of gDNA derived from transformants. The transcript level and expression amount of recombinant hIFN-γ protein were evaluated in various events using RT-PCR and enzyme-linked immunosorbent assay (ELISA) techniques. A disagreement between the transcript level and the amount of recombinant protein with an inverse correlation between transgene copy number and expression level observed in some events, probably showing translational gene silencing and co-suppression or silencing, respectively. These results were also compared with segregation ratios of hygromycin-resistant phenotype in T1 plants of each line and found to be, in general, consistent.

在转基因植物中,转基因拷贝数会严重影响转基因的表达水平和遗传稳定性。因此,表征转基因植物的第一步就是估算整合到植物基因组中的转基因拷贝数。我们采用定量实时 PCR(qRT-PCR)技术,以 axi1 基因为内源对照,测定了人干扰素-γ(hIFN-γ)和百菌清磷酸转移酶 II(hptII)转基因在 18 个转基因烟草品系 T0 代基因组中的拷贝数。通过优化 PCR 条件,我们对 T0 转化株中的 1、2、3 和/或 4 个转基因拷贝数进行了高度精确的估计。此外,对 hIFN-γ 转基因和 hptII 选择性标记基因拷贝数的估计表明,T-DNA 的重排在转基因烟草中经常发生。通过对转化株的 gDNA 进行 Southern 印迹分析,还估算出了转基因的拷贝数。利用 RT-PCR 和酶联免疫吸附试验(ELISA)技术评估了重组 hIFN-γ 蛋白在不同事件中的转录水平和表达量。在某些事件中,转录本水平与重组蛋白量之间存在差异,转基因拷贝数与表达水平之间呈反相关,这可能分别显示了转译基因沉默和共抑制或沉默。这些结果还与各品系 T1 植株的抗潮霉素表型的分离比进行了比较,发现两者基本一致。
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引用次数: 0
Effect of phosphorus deficiency on the expression and protein localization of ENHANCER OF TRY AND CPC3 gene in Arabidopsis roots 缺磷对拟南芥根中 TRY 和 CPC3 基因表达及蛋白定位的影响
IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-01 DOI: 10.1007/s13562-024-00880-6
Juri Wakamatsu, Jun Hosokawa, Rumi Tominaga

Increased hair in Arabidopsis roots during phosphorus deficiency is an important adaptation mechanism. The CAPRICE (CPC) gene, along with its six homologs, plays a pivotal role in root hair formation. Our previous study indicated upregulated expression of ENHANCER OF TRY AND CPC1 (ETC1) and ENHANCER OF TRY AND CPC3 (ETC3) homologous genes in response to phosphorus deficiency. ETC1 translocates from the root sub-epidermis to epidermal cells, thereby increasing the number of root hairs. In this study, we specifically focused on ETC3 to explore its function in this process. Our results showed that etc1, etc3, and etc1 etc3 mutants did not induce as much root hairs in response to phosphorus deficiency as the wild type. The upregulation of ETC1 and ETC3 in response to phosphorus deficiency was restricted to the roots, with no such response observed in leaves. Furthermore, examination of ETC3:GFP protein localization revealed that ETC3 expression was induced in the internal root tissues under phosphorus deficiency, without migrating to epidermal cells as observed for ETC1. This finding suggests that ETC3 may indirectly contribute to root hair formation through a different mechanism. Therefore, elucidating the complex process that governs root hair increase in phosphorus-deficient plants can lead to future advances in plant breeding and nutrient stress adaptation.

拟南芥在缺磷期间根毛增多是一种重要的适应机制。CAPRICE(CPC)基因及其六个同源基因在根毛形成过程中起着关键作用。我们之前的研究表明,TRY 和 CPC1 的 ENHANCER(ETC1)和 TRY 和 CPC3 的 ENHANCER(ETC3)同源基因在缺磷时表达上调。ETC1 从根的表皮下细胞转位到表皮细胞,从而增加根毛的数量。在本研究中,我们特别关注了 ETC3,以探索其在这一过程中的功能。结果表明,etc1、etc3和etc1 etc3突变体对缺磷的诱导根毛数量不如野生型。ETC1 和 ETC3 对缺磷反应的上调仅限于根部,在叶片中没有观察到这种反应。此外,对 ETC3:GFP 蛋白定位的研究发现,在缺磷情况下,ETC3 在根内部组织中被诱导表达,而没有像 ETC1 那样迁移到表皮细胞。这一发现表明,ETC3 可能通过不同的机制间接促进了根毛的形成。因此,阐明缺磷植物根毛增加的复杂过程可促进未来植物育种和营养胁迫适应的进步。
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Journal of Plant Biochemistry and Biotechnology
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