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| 1 | A method for the production and expedient screening of CRISPR/Cas9-mediated non-transgenic mutant plants显示文摘Developing CRISPR/Cas9-mediated non-transgenic mutants in asexually propagated perennial crop plants is challenging but highly desirable.Here,we report a highly useful method using an Agrobacterium-mediated transient CRISPR/Cas9 gene expression system to create non-transgenic mutant plants without the need for sexual segregation.We have also developed a rapid,cost-effective,and high-throughput mutant screening protocol based on Illumina sequencing followed by high-resolution melting(HRM)analysis.Using tetraploid tobacco as a model species and the phytoene desaturase(PDS)gene as a target,we successfully created and expediently identified mutant plants,which were verified as tetra-allelic mutants.We produced pds mutant shoots at a rate of 47.5%from tobacco leaf explants,without the use of antibiotic selection.Among these pds plants,17.2%were confirmed to be non-transgenic,for an overall non-transgenic mutation rate of 8.2%.Our method is reliable and effective in creating non-transgenic mutant plants without the need to segregate out transgenes through sexual reproduction.This method should be applicable to many economically important,heterozygous,perennial crop species that are more difficult to regenerate. | Longzheng Chen Wei Li Lorenzo Katin-Grazzini Jing Ding Xianbin Gu Yanjun Li Tingting Gu Ren Wang Xinchun Lin Ziniu Deng Richard J.McAvoy Frederick G.Gmitter Jr. Zhanao Deng Yunde Zhao Yi Li | 2018 | Horticulture Research2018，5，1： | 12 |
| 2 | Growth performance and physiological parameters of the second generation selected and control groups of Pinctada martensii显示文摘In May of 2007,the second generation selected (SS) and control (SC) groups of pearl oyster Pinctada martensii were established by selecting 10% breeders with the largest and mean shell length,respectively,from the first generation selected group.Growth performance of the SS and SC groups were compared on the basis of measurement data at Days 8,18,60,95,195 and 365.On Day 365,100 individuals (60.0–75.0 mm at shell length) were sampled from each group and then subjected to the experiment where physiological parameters such as filtrate rate,oxygen consumption and ammonia excretion were measured at 15,20,25 and 30°C.The results show that the SS group had significantly larger mean shell length and shell height than the SC group at Days 8,18,60,95,195 and 365 (P <0.05).The genetic gains at different ages varied from 6.0% to 17.0% for shell length and 5.7% to 14.6% for shell height,respectively.At 15,20,25 and 30 ° C,the SS groups had significantly larger filtrate rate than the SC group (P <0.05).At 15 and 25 °C,the differences in oxygen consumption rate between the SS and SC groups were not significant (P >0.05).At 20 and 30 °C,however,the oxygen consumption rate of the SS group was significantly larger than the SC group (P <0.05).At 15,20,25 and 30 °C,there were no significant differences in ammonia excretion rate between the SS and SC groups (P >0.05).The present results indicate that there existed considerable genetic variability in the base population and a further selection could be likely fruitful.Mass selection for faster growth might produce animals that had higher intake of metabolizable energy by virtue of faster filtrating behavior. | DENG Yuewen YU Ziniu DU Xiaodong WANG Qingheng FU Shao | 2011 | Acta Oceanologica Sinica2011，30，2： | 10 |
| 3 | Citron C-05 inhibits both the penetration and colonization of Xanthomonas citri subsp.citri to achieve resistance to citrus canker disease显示文摘Citrus canker,caused by Xanthomonas citri subsp.citri(Xcc),is a serious bacterial disease that affects citrus production worldwide.Citron C-05(Citrus medica)is the only germplasm in the Citrus genus that has been identified to exhibit strong resistance to Xcc.However,it has not been determined when,where,and how Xcc is restricted in the tissues of Citron C-05 during the infection process.In the present study,we investigated the spatiotemporal growth dynamics of an eGFP-labeled virulent Xcc(eGFP-Xcc)strain in Citron C-05 along with five susceptible biotypes(i.e.,lemon,pummelo,sour orange,sweet orange,and ponkan mandarin)upon inoculation via the spraying or leaf infiltration of a bacterial suspension.The results from extensive confocal laser scanning microscopy analyses showed that while Xcc grew rapidly in plants of all five susceptible genotypes,Xcc was severely restricted in the epidermal and mesophyll cell layers of the leaves of Citron C-05 in the early stage of infection.Not surprisingly,resistance against Xcc in Citron C-05 was found to be associated with the production of reactive oxygen species and hypersensitive response-like cell death,as well as greater upregulation of several defense-related genes,including a pathogenesis-related gene(PR1)and a glutathione S-transferase gene(GST1),compared with sweet orange as a susceptible control.Taken together,our results not only provide further valuable details of the spatiotemporal dynamics of the host entry,propagation,and spread of Xcc in both resistant and susceptible citrus plants but also suggest that resistance to Xcc in Citron C-05 may be attributed to the activation of multiple defense mechanisms. | Hongyan Fu Mingming Zhao Jing Xu Limei Tan Jian Han Dazhi Li Meijun Wang Shunyuan Xiao Xianfeng Ma Ziniu Deng | 2020 | Horticulture Research2020，7，1： | 2 |
| 4 | Early embryonic transcriptomes of Zeugodacus tau provide insight into sex determination and differentiation genes显示文摘Zeugodacus tau(Walker)is an invasive pest.The sterile insect technique is an environment-friendly method for pest control.Understanding the mechanism of sex determination will contribute to improving efficiency of this technique.In this study,we identified the transformer(tra)gene in Z.tau.One female-specific and two male-specific isoforms of tra were found in Z.tau,and the male-specific splicing pattern of tra was found to occur 5 h after egg laying.We performed transcriptome sequencing at 1 h(E1),5 h(E5),and 9 h(E9)after egg laying and obtained high-quality transcriptome libraries of early embryo development.We identified 13297 and 11713 differentially expressed genes(DEGs)from E5 versus E1 and E9 versus E1 comparisons,respectively.To explore the potential functions of the DEGs during embryonic development,Gene Ontology,Clusters of Orthologous Groups of proteins,and Kyoto Encyclopedia of Genes and Genomes analyses were performed.Twenty-six genes potentially involved in sex determination or differentiation,including Maleness-on-the-Y(MoY),were identified in Z.tau.To verify the transcriptome results,15 genes were selected for quantitative real-time PCR validation.The results were consistent with the transcriptome sequencing results.Moreover,U2 small nuclear riboprotein auxiliary factor(U2AF-50),female lethal d(fl(2)d),and virilizer(vir)were highly expressed at E5,indicating that they may be related to the sex-specific splicing of tra.Further functional analysis is needed to confirm this speculation.Our data provide an insight into the mechanism underlying sex determination and differentiation in tephritid species. | Peipei Liu Wenping Zheng Jiao Qiao Ziniu Li Zhurong Deng Yimin Yuan Hongyu Zhang | 2022 | Insect Science2022，29，3： | 1 |
| 5 | Endogenous auxin and its manipulation influence in vitro shoot organogenesis of citrus epicotyl explants显示文摘Endogenous auxin is an important regulator of in vivo organ development,but its role in in vitro organogenesis is unclear.It has been observed that the basal end of epicotyl cuttings of juvenile citrus seedlings produces fewer shoots than the apical end.Here,we report that elevated endogenous auxin levels in the basal end of citrus epicotyl cuttings are inhibitory for in vitro shoot organogenesis.Using transgenic citrus plants expressing an auxin-inducible GUS reporter gene,we have observed elevated levels of auxin at the basal end of stem cuttings that are mediated by polar auxin transport.Depleting endogenous auxin or blocking polar auxin transport enhances shoot organogenesis.An auxin transport inhibitor,N-1-naphthylphthalamic acid(NPA),can also enhance shoot organogenesis independent of its action on polar auxin transport.Finally,we demonstrate that the promotional effects of depleting endogenous auxin or blocking polar auxin transport on shoot organogenesis are cytokinin-dependent.Our study thus provides meaningful insights into possible roles of endogenous auxin and polar auxin transport,as well as auxin–cytokinin interactions,in in vitro shoot organogenesis.Meanwhile,our results may also provide practical strategies for improving in vitro shoot organogenesis for citrus and many other plant species. | Wei Hu Sabrina Fagundez Lorenzo Katin-Grazzini Yanjun Li Wei Li Yingnan Chen Xiaomin Wang Ziniu Deng Shenxi Xie Richard J McAvoy Yi Li | 2017 | Horticulture Research2017，4，1： | 1 |
| 6 | Defence-related gene expression in transgenic lemon plants producing an antimicrobial Trichoderma harzianum endochitinase during fungal infection显示文摘 | Gaetano Distefano Stefano La Malfa Alessandro Vitale Matteo Lorito Ziniu Deng Alessandra Gentile | 2008 | Transgenic Research2008，，5： | 1 |
| 7 | Cloning of a novel glutathione S-transferase 3 ( GST3 ) gene and expressionanalysis in pearl oyster, Pinctada martensii显示文摘 | Jinhui Chen Shu Xiao Yuewen Deng Xiaodong Du Ziniu Yu | 2011 | Fish and Shellfish Immunology2011，，6： | 1 |
| 8 | Transformation of sweet orange [Citrus sinensis (L.) Osbeck] with pthA-nls for acquiring resistance to citrus canker disease显示文摘 | Li Yang Chunhua Hu Na Li Jiayin Zhang Jiawen Yan Ziniu Deng | 2011 | 2011 (1-2)2011，，1： | 1 |
| 9 | Bioinformatics and Expression Analysis of CPMAX2 in Citrange显示文摘Transgenic citrange lines with rol ABC genes behave rosette branching and extreme dwarfing.To explore the regulatory mechanism of plant hormones in axillary shoot growth of transgenic citrange,a full length cD NA of CPMAX2 was cloned from citrange [Citrus sinensis(L.) Osb × Poncirus trifoliate(L.) Raf.] by RT-PCR in this study.The expression of CPMAX2 was detected in axillary tender leaves of 3 transgenic citrange lines and the wild type.Additionally,we constructed an over-expression vector CPMAX2-p CAMBIA1301 for further study.The results showed that the c DNA sequence and its putative peptide sequence shared 99.66% and 99.14% of identity with its Citrus sinensis ortholog MAX2.The deduced amino acid sequence contained putatively one F-box and two LRR repeat domains that are highly conserved in MAX2 genes.CPMAX2 was obviously down-expressed in tender leaves of 3 transgenic citrange lines compared with the wild type.The results suggest CPMAX2 play an important role in regulating the rosette shoot growth in transgenic citrange with rol ABC genes. | YUAN Feirong YUAN Yaoming LI Zhixin Alessandra GENTILE DENG Ziniu | 2016 | Horticultural Plant Journal2016，2，3： | 0 |
| 10 | Comparative Genomics Reveals Evolutionary Drivers of Sessile Life and Left-right Shell Asymmetry in Bivalves显示文摘Bivalves are species-rich mollusks with prominent protective roles in coastal ecosystems.Across these ancient lineages,colony-founding larvae anchor themselves either by byssus production or by cemented attachment.The latter mode of sessile life is strongly molded by left-right shell asymmetry during larval development of Ostreoida oysters such as Crassostrea hongkongensis.Here,we sequenced the genome of C.hongkongensis in high resolution and compared it to reference bivalve genomes to unveil genomic determinants driving cemented attachment and shell asymmetry.Importantly,loss of the homeobox gene Antennapedia(Antp)and broad expansion of lineagespecific extracellular gene families are implicated in a shift from byssal to cemented attachment in bivalves.Comparative transcriptomic analysis shows a conspicuous divergence between leftright asymmetrical C.hongkongensis and symmetrical Pinctada fucata in their expression profiles.Especially,a couple of orthologous transcription factor genes and lineage-specific shell-related gene families including that encoding tyrosinases are elevated,and may cooperatively govern asymmetrical shell formation in Ostreoida oysters. | Yang Zhang Fan Mao Shu Xiao Haiyan Yu Zhiming Xiang Fei Xu Jun Li Lili Wang Yuanyan Xiong Mengqiu Chen Yongbo Bao Yuewen Deng Quan Huo Lvping Zhang Wenguang Liu Xuming Li Haitao Ma Yuehuan Zhang Xiyu Mu Min Liu Hongkun Zheng Nai-Kei Wong Ziniu Yu | 2022 | Genomics, Proteomics & Bioinformatics2022，20，6： | 0 |
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