CRISPR-Cas endonucleases mediate prokaryotic adaptive immunity by targeting foreign nucleic acids.CRISPR/Cas13b is a class 2 type VI-B ribonuclease that targets and cleaves single-stranded RNA.It exhibits higher RNA i...CRISPR-Cas endonucleases mediate prokaryotic adaptive immunity by targeting foreign nucleic acids.CRISPR/Cas13b is a class 2 type VI-B ribonuclease that targets and cleaves single-stranded RNA.It exhibits higher RNA interference activity than Cas13a and Cas13c and causes fewer collateral effects than RxCas13d in mammalian cells.However,a programmable CRISPR/Cas13b-mediated RNA interference system for endogenous transcripts in rice has not yet been established.Here,we developed a CRISPR/Cas13b-mediated system to target endogenous transcripts in rice.Our CRISPR/Cas13b system could inhibit multiple endogenous mRNAs simultaneously.In addition,this system efficiently repressed endogenous long noncoding RNAs with more than 50% inhibition in stable transgenic plants.Furthermore,we found only weak collateral effects of the CRISPR/Cas13b-mediated system at the transcriptome-wide level,and no difference in the agronomic traits of stable transgenic rice in the field.We present a programmable CRISPR/Cas13b-mediated knockdown system for rice,offering a potential biotechnological tool for functional genomics and crop improvement.展开更多
This research paper addresses,from an ecogeographic perspective within a localised context,a new concept of nature conservation within modern farming systems:the direct seeding technique based on the principles of the...This research paper addresses,from an ecogeographic perspective within a localised context,a new concept of nature conservation within modern farming systems:the direct seeding technique based on the principles of the Conservation Agriculture System(CAS).The adoption of CAS aims to increase soil fertility,promote biodiversity,and sustain production,making it one of the most effective adaptation solutions available to address the challenges of climate change.CAS is defined as a farming system based on three key principles:minimal soil disturbance(reduced or zero tillage),maintaining a permanent soil cover(with residues from previous crops),and adopting crop rotation(diversifying crops rather than limiting them to a single type in consecutive seasons).However,there is limited research and a lack of scientific studies on the implementation of conservation agriculture in developing nations such as Morocco,particularly in the Had Kourt region.This study aims to assess the feasibility of CAS compared to traditional agricultural systems by surveying the opinions of farmers who have experience with both systems and comparing the outcomes of their practices based on simple indicators as an initial stage,with further analysis of additional indicators planned through subsequent scientific investigations.The sampling method used in this study is non-discriminatory,as semi-structured interviews were conducted with a group of farmers to gather their opinions on CAS.The adoption of conservation agriculture has been linked to prior knowledge of the system,acquired through training and fieldwork via application platforms,which remain limited in scope.展开更多
非特异性脂质转移蛋白(non-specific lipid transfer protein,nsLTPs)广泛存在于植物界中,参与许多关键的生物学过程,如花粉的发育、种子发育及细胞壁的延伸等。OsLTPL166基因编码区长度是465 bp,其编码的蛋白质由154个氨基酸组成;根据...非特异性脂质转移蛋白(non-specific lipid transfer protein,nsLTPs)广泛存在于植物界中,参与许多关键的生物学过程,如花粉的发育、种子发育及细胞壁的延伸等。OsLTPL166基因编码区长度是465 bp,其编码的蛋白质由154个氨基酸组成;根据跨膜结构域及信号肽预测,OsLTPL166在13~35氨基酸处含有跨膜结构域,N端存在一个32个氨基酸残基的信号肽,切割位点位于氨基酸残基32和33之间,产生具有122个氨基酸残基的成熟蛋白质。组织表达分析发现,OsLTPL166仅在种子中特异性表达,且种子发育后期表达量较高。为了研究水稻Os LTPL166在种子发育中的功能,本研究以‘浙辐粳83’为遗传背景构建了敲除载体,利用CRISPR/Cas9技术对OsLTPL166进行定点编辑,共获得24个独立的转基因株系,其中纯合突变株系有7株,编辑方式主要为碱基的插入和缺失,导致氨基酸序列发生移码,成功获得OsLTPL166功能缺陷型突变体。本研究为进一步探究该基因在种子发育进程中的生物学功能提供了遗传材料。展开更多
基金supported by the Natural Science Foundation of Zhejiang Province,China(Grant Nos.LZ22C150002 and LR24C150001)the National Key Research and Development Program of China(Grant Nos.2021YFF1000402 and 2022YFD1401600)+1 种基金the National Natural Science Foundation of China(Grant No.32170262)the Fundamental Research Funds for the Central Universities,China(Grant No.K20240124).
文摘CRISPR-Cas endonucleases mediate prokaryotic adaptive immunity by targeting foreign nucleic acids.CRISPR/Cas13b is a class 2 type VI-B ribonuclease that targets and cleaves single-stranded RNA.It exhibits higher RNA interference activity than Cas13a and Cas13c and causes fewer collateral effects than RxCas13d in mammalian cells.However,a programmable CRISPR/Cas13b-mediated RNA interference system for endogenous transcripts in rice has not yet been established.Here,we developed a CRISPR/Cas13b-mediated system to target endogenous transcripts in rice.Our CRISPR/Cas13b system could inhibit multiple endogenous mRNAs simultaneously.In addition,this system efficiently repressed endogenous long noncoding RNAs with more than 50% inhibition in stable transgenic plants.Furthermore,we found only weak collateral effects of the CRISPR/Cas13b-mediated system at the transcriptome-wide level,and no difference in the agronomic traits of stable transgenic rice in the field.We present a programmable CRISPR/Cas13b-mediated knockdown system for rice,offering a potential biotechnological tool for functional genomics and crop improvement.
文摘This research paper addresses,from an ecogeographic perspective within a localised context,a new concept of nature conservation within modern farming systems:the direct seeding technique based on the principles of the Conservation Agriculture System(CAS).The adoption of CAS aims to increase soil fertility,promote biodiversity,and sustain production,making it one of the most effective adaptation solutions available to address the challenges of climate change.CAS is defined as a farming system based on three key principles:minimal soil disturbance(reduced or zero tillage),maintaining a permanent soil cover(with residues from previous crops),and adopting crop rotation(diversifying crops rather than limiting them to a single type in consecutive seasons).However,there is limited research and a lack of scientific studies on the implementation of conservation agriculture in developing nations such as Morocco,particularly in the Had Kourt region.This study aims to assess the feasibility of CAS compared to traditional agricultural systems by surveying the opinions of farmers who have experience with both systems and comparing the outcomes of their practices based on simple indicators as an initial stage,with further analysis of additional indicators planned through subsequent scientific investigations.The sampling method used in this study is non-discriminatory,as semi-structured interviews were conducted with a group of farmers to gather their opinions on CAS.The adoption of conservation agriculture has been linked to prior knowledge of the system,acquired through training and fieldwork via application platforms,which remain limited in scope.
