aBIOTECH

Tissues and organs within a living organism are coordinated, but the underlying mechanisms are not well understood. The shoot apical meristem (SAM) continually produces lateral organs, such as leaves, from its peripheral zone. Because of their close proximity, SAM and lateral organs interact during plant development. Existing lateral organs influence the positions of newly formed organs to determine the phyllotaxis. The SAM not only produces lateral organs, but also influences their morphogenesis. In particular, the SAM promotes leaf polarity determination and leaf blade formation. Furthermore, lateral organs help the SAM to maintain homeostasis by restricting stem cell activity. Recent advances have started to elucidate how SAM and lateral organs patterning and growth are coordinated in the shoot apex. In this review, we discuss recent findings on the interaction between SAM and lateral organs during plant development. In particular, polar auxin transport appears to be a commonly used coordination mechanism.

Chunmei Guan

Yuling Jiao

Interplay between the shoot apical meristem and lateral organs

茎尖分生组织与侧生器官的相互作用

Abstract As a conserved epigenetic mark, DNA cytosine methylation, at the 5’ position (5-mC), plays important roles in multiple biological processes, including plant immunity. However, the involvement of DNA methylation in the determinants of virulence of phytopathogenic fungi remains elusive. In this study, we profiled the DNA methylation patterns of the phytopathogenic fungus Verticillium dahliae , one of the major causal pathogens of Verticillium wilt disease that causes great losses in many crops, and explored its contribution in fungal pathogenicity. We reveal that DNA methylation modification is present in V. dahliae and is required for its full virulence in host plants. The major enzymes responsible for the establishment of DNA methylation in V. dahliae were identified. We provided evidence that DNA methyltransferase-mediated establishment of DNA methylation pattern positively regulates fungal virulence, mainly through repressing a conserved protein kinase VdRim15-mediated Ca 2+ signaling and ROS production, which is essential for the penetration activity of V. dahliae . In addition, we further demonstrated that histone H3 lysine 9 trimethylation (H3K9me3), another heterochromatin marker that is closely associated with 5-mC in eukaryotes, also participates in the regulation of V. dahliae pathogenicity, through a similar mechanism. More importantly, DNA methyltransferase genes VdRid , VdDnmt5, as well as H3K9me3 methyltransferase genes, were greatly induced during the early infection phase, implying that a dynamic regulation of 5-mC and H3K9me3 homeostasis is required for an efficient infection. Collectively, our findings uncover an epigenetic mechanism in the regulation of phytopathogenic fungal virulence.

Yun-Ya Chen

Chen Zhu

Jian-Hua Zhao

Ting Liu

Feng Gao

Ying-Chao Zhang

Cheng-Guo Duan

DNA methylation-dependent epigenetic regulation of Verticillium dahliae virulence in plants

植物中Verticillium dahliae毒力的DNA甲基化依赖性表观遗传调控

Flowering links vegetative growth and reproductive growth and involves the coordination of local environmental cues and plant genetic information. Appropriate timing of floral initiation and maturation in both wild and cultivated plants is important to their fitness and productivity in a given growth environment. The domestication of plants into crops, and later crop expansion and improvement, has often involved selection for early flowering. In this review, we analyze the basic rules for photoperiodic adaptation in several economically important and/or well-researched crop species. The ancestors of rice (Oryza sativa), maize (Zea mays), soybean (Glycine max), and tomato (Solanum lycopersicum) are short-day plants whose photosensitivity was reduced or lost during domestication and expansion to high-latitude areas. Wheat (Triticum aestivum) and barley (Hordeum vulgare) are long-day crops whose photosensitivity is influenced by both latitude and vernalization type. Here, we summarize recent studies about where these crops were domesticated, how they adapted to photoperiodic conditions as their growing area expanded from domestication locations to modern cultivating regions, and how allelic variants of photoperiodic flowering genes were selected during this process. A deeper understanding of photoperiodic flowering in each crop will enable better molecular design and breeding of high-yielding cultivars suited to particular local environments.The online version contains supplementary material available at 10.1007/s42994-021-00039-0.

Xiaoya Lin

Chao Fang

Baohui Liu

Fanjiang Kong

Natural variation and artificial selection of photoperiodic flowering genes and their applications in crop adaptation

光周期开花基因的自然变异与人工选择及其在作物适应性中的应用

The CRISPR-Cas genome editing tools are revolutionizing agriculture and basic biology with their simplicity and precision ability to modify target genomic loci. Software-predicted guide RNAs (gRNAs) often fail to induce efficient cleavage at target loci. Many target loci are inaccessible due to complex chromatin structure. Currently, there is no suitable tool available to predict the architecture of genomic target sites and their accessibility. Hence, significant time and resources are spent on performing editing experiments with inefficient guides. Although in vitro-cleavage assay could provide a rough assessment of gRNA efficiency, it largely excludes the interference of native genomic context. Transient in-vivo testing gives a proper assessment of the cleavage ability of editing reagents in a native genomic context. Here, we developed a modified protocol that offers highly efficient protoplast isolation from rice,Arabidopsis,and chickpea, using a sucrose gradient, transfection using PEG (polyethylene glycol), and validation of single guide RNAs (sgRNAs) cleavage efficiency of CRISPR-Cas9. We have optimized various parameters for PEG-mediated protoplast transfection and achieved high transfection efficiency using our protocol in both monocots and dicots. We introduced plasmid vectors containing Cas9 and sgRNAs targeting genes in rice,Arabidopsis,and chickpea protoplasts. Using dual sgRNAs, our CRISPR-deletion strategy offers straightforward detection of genome editing success by simple agarose gel electrophoresis. Sanger sequencing of PCR products confirmed the editing efficiency of specific sgRNAs. Notably, we demonstrated that isolated protoplasts can be stored for up to 24/48 h with little loss of viability, allowing a pause between isolation and transfection. This high-efficiency protocol for protoplast isolation and transfection enables rapid (less than 7 days) validation of sgRNA cleavage efficiency before proceeding with stable transformation. The isolation and transfection method can also be utilized for rapid validation of editing strategies, evaluating diverse editing reagents, regenerating plants from transfected protoplasts, gene expression studies, protein localization and functional analysis, and other applications.

Debasmita Panda

Subhasis Karmakar

Manaswini Dash

Swagat Kumar Tripathy

Priya Das

Sagar Banerjee

Yiping Qi

Sanghamitra Samantaray

Pradipta Kumar Mohapatra

Mirza J. Baig

Kutubuddin A. Molla

Optimized protoplast isolation and transfection with a breakpoint: accelerating Cas9/sgRNA cleavage efficiency validation in monocot and dicot

优化原生质体分离与转染：在单子叶和双子叶植物中加速Cas9/sgRNA切割效率验证的关键点

Genome editing holds great promise for the molecular breeding of plants, yet its application is hindered by the shortage of simple and effective means of delivering genome editing reagents into plants. Conventional plant transformation-based methods for delivery of genome editing reagents into plants often involve prolonged tissue culture, a labor-intensive and technically challenging process for many elite crop cultivars. In this review, we describe various virus-based methods that have been employed to deliver genome editing reagents, including components of the CRISPR/Cas machinery and donor DNA for precision editing in plants. We update the progress in these methods with recent successful examples of genome editing achieved through virus-based delivery in different plant species, highlight the advantages and limitations of these delivery approaches, and discuss the remaining challenges.

Yilin Shen

Tao Ye

Zihan Li

Torotwa Herman Kimutai

Hao Song

Xiaoou Dong

Jianmin Wan

Exploiting viral vectors to deliver genome editing reagents in plants

利用病毒载体在植物中传递基因编辑试剂

Abstract Crop breeding requires a balance of tradeoffs among key agronomic traits caused by gene pleiotropy. The molecular manipulation of genes can effectively improve target traits, but this may not reduce gene pleiotropy, potentially leading to undesirable traits or even lethal conditions. However, molecular editing of cis -regulatory elements (CREs) of target genes may facilitate the dissection of gene pleiotropy to fine-tune gene expression. In this study, we developed a pipeline, in potato, which employs open chromatin to predict candidate CREs, along with both transient and genetic assays to validate the function of CREs and CRISPR/Cas9 to edit candidate CREs. We used StCDF1 as an example, a key gene for potato tuberization and identified a 288 bp-core promoter region, which showed photoperiodic inducibility. A homozygous CRISPR/Cas9-editing line was established, with two deletions in the core promoter, which displayed a reduced expression level, resulting in late tuberization under both long-day and short-day conditions. This pipeline provides an alternative pathway to improve a specific trait with limited downside on other phenotypes.

Min Wan

Handan Xie

Hongwei Guo

Shenglin Jing

Deying Zeng

Bing Li

Bo Zhu

Zixian Zeng

Developing a pipeline for identification, characterization and molecular editing of cis-regulatory elements: a case study in potato

开发顺式调控元件的鉴定、表征和分子编辑流程：以马铃薯为例

Abstract Tomato ( Solanum lycopersicum ) and potato ( Solanum tuberosum ), two integral crops within the nightshade family, are crucial sources of nutrients and serve as staple foods worldwide. Molecular genetic studies have significantly advanced our understanding of their domestication, evolution, and the establishment of key agronomic traits. Recent studies have revealed that epigenetic modifications act as “molecular switches”, crucially regulating phenotypic variations essential for traits such as fruit ripening in tomatoes and tuberization in potatoes. This review summarizes the latest findings on the regulatory mechanisms of epigenetic modifications in these crops and discusses the integration of biotechnology and epigenomics to enhance breeding strategies. By highlighting the role of epigenetic control in augmenting crop yield and adaptation, we underscores its potential to address the challenges posed by a growing global population as well as changing climate.

Pingxian Zhang

Yuehui He

Sanwen Huang

Unlocking epigenetic breeding potential in tomato and potato

解锁番茄和马铃薯的表观遗传育种潜力

Abstract The effect of fungicides on the plant-rhizosphere microbiome is a subject of ongoing debate, but whether any alteration in the rhizosphere microbiome could affect plant health is an issue that has not been thoroughly investigated. To address this deficiency, we analyzed the rhizosphere microbiome of wilt disease—resistant and disease-susceptible cucumber cultivars to determine whether (and which) plant-associated microorganisms have a role in disease resistance. We further assessed whether the fungicides thiophanate-methyl and carbendazim affect the rhizosphere microbiome, which may contribute to the plant’s immune response. Based on results acquired with both radicle-inoculation and soil-inoculation methods, cultivars Longyuanxiuchun (LYXC) and Shuyan2 (SY2) were identified as being disease resistant, whereas Zhongnong6 (ZN6) and Zhongnong38 (ZN38) were susceptible. The microbiome structure differed substantially between the resistant and susceptible plants, with LYXC and SY2 each having a significantly greater Shannon index than Zhongnong38. These results revealed that the disease-resistant cucumber cultivars recruited more beneficial bacteria, i.e., Bacillus , in their rhizosphere soil; as such, Bacillus was identified as a keystone genus in the microbial co-occurrence network. Thus, the presence of Bacillus may help cucumbers defend against fungal pathogens within the rhizosphere. Bacillus subtilis strain LD15, which was isolated from LYXC rhizosphere soil, could suppress pathogen growth, in vitro, and reduce disease severity in pot assays. Moreover, evidence also confirmed the accumulation of LD1 in the rhizosphere soil of resistant cucumber cultivars. For LYXC, application of thiophanate-methyl or carbendazim altered the microbiome structure, decreased bacterial diversity, and reduced the abundance of Bacillus species. Finally, pot assays verified that fungicide application decreased the proportion of LD15 in rhizosphere soil. From a microbial perspective, thiophanate-methyl and carbendazim may weaken the rhizobacteria-mediated defense response of cucumbers against cucumber Fusarium wilt disease. Our findings reveal a role for the rhizosphere microbiome in protecting plants from pathogens and constitute a reference for assessing the ecotoxicological risk of pesticides to non-target soil microorganisms.

Kai Cui

Xiaoming Xia

Youwei Wang

Yueli Zhang

Ying Zhang

Junli Cao

Jun Xu

Fengshou Dong

Xingang Liu

Xinglu Pan

Yongquan Zheng

Xiaohu Wu

Thiophanate-methyl and its major metabolite carbendazim weaken rhizobacteria-mediated defense responses in cucumbers against Fusarium wilt

甲基硫菌灵及其主要代谢物多菌灵削弱黄瓜根际细菌介导的抗枯萎病防御反应

Abstract The barley genome encodes a complete set of MADS-box proteins sharing homology with components of the ABCDE model, which explains the molecular basis of floral organ identity in angiosperm flowers. Although the E-class members are universally expressed across floral whorls and crucial for flower development in Arabidopsis and rice, the functional role of the barley E-class LOFSEP subfamily (comprising MADS1, MADS5, and MADS34) remains elusive, particularly during spikelet formation. Here, we characterize the single, double and triple lofsep mutants in barley in an attempt to overcome the anticipated genetic redundancy. Surprisingly, loss of function of all LOFSEP members only disturbs lemma development, either converting this hull organ into a leaf-like structure or reducing its size. The inner organs, including lodicules, anthers and pistil remain unaffected. A systematic interrogation of how ABCDE class genes are affected in all whorls of the mutants was undertaken. Generally, in the lemma and palea of the lofsep mutants, A- and E-class genes are hyperactivated, B- and C- classes are slightly repressed, and D-class genes show unchanged expression in these inner organs. Intriguingly, loss of function of MADS6 , an AGL6 member closely related to the E-class genes, leads to most organs being transformed into lemma-like organs with new spikelets generated from the center of the flower. Contrasting with rice, these findings suggest barley LOFSEPs may have regressed in determining floral organ identity, and this could be partially compensated by HvMADS6.

Chaoqun Shen

Xiujuan Yang

Duoxiang Wang

Gang Li

Matthew R. Tucker

Functional retrogression of LOFSEPs in specifying floral organs in barley

大麦中LOFSEPs在指定花器官功能退化

Abstract Viable pollen is crucial for fertilization, but pollen is generally highly susceptible to heat stress. A quick, reliable method for testing the heat-stress tolerance of pollen is needed to improve the heat-stress tolerance in plants, but current methods require considerable space and labor. In addition, many such methods only test tolerance to a single constant temperature, making it time-consuming to screen heat tolerance over a wide temperature range and to examine the dynamics of pollen viability at different temperatures. To address this issue, we aimed to: (1) develop an easy, reliable method for measuring pollen viability at different temperatures; and (2) identify the best temperature range for screening pollen with high heat-stress tolerance. We harvested mature pollen from wheat ( Triticum aestivum ) plants and transferred it to a 96-well plate filled with liquid medium containing sucrose. We placed the plate in a PCR machine operating under a gradient PCR program to simultaneously test a range of temperatures. After incubating the pollen for 4 h, at temperatures ranging from 21.9 to 47 °C, we examined the pollen grains under a light microscope and employed a specific image analysis pipeline to assess the effects of temperature on pollen morphology, germination, and tube growth. This method facilitated the high-throughput screening of many pollen samples, enabling rapid, reliable, and precise analysis of pollen viability in response to temperature. Our approach should be applicable to other plant species and could be used to identify quantitative trait loci or genes influencing heat stress tolerance in pollen for breeding programs.

短名	Abiotech.
Journal Impact	4.70
国际分区	PLANT SCIENCES(Q1)
期刊索引	中科院 4 区中国科学引文数据库

ISSN	2096-6326, 2662-1738
h-index	16
国内分区	农林科学(4区)农林科学生物工程与应用微生物(4区)农林科学植物科学(4区)

出版信息	出版商: Springer，出版周期: ，期刊类型: journal，开源期刊: 否
基本数据	创刊年份: 2020，原创研究文献占比: 47.83%，自引率:， Gold OA占比: 33.33%

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