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Generation of selectable marker-free soft transgenic rice with transparent kernels by downregulation of SSSⅡ-2

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The Crop Journal 2020年第1期8卷 53-61页

作者： Zhihao Xu Meimei Yu Youwei Yin Cenwen Zhu Wen Ji Changquan Zhang Qianfeng Li Honggen Zhang Shuzu Tang Hengxiu Yu Qiaoquan Liu Jiangsu Key laboratory of Crop genetics and Physiology Key Laboratory of Plant Functional Genomics of the Ministry of EducationJiangsu Key Laboratory of Crop Genomics and Molecular BreedingJiangsu Co-Innovation Center for Modern Production Technology of Grain CropsCollege of AgricultureYangzhou University

The amylose content(AC) of rice endosperm starch varies from 0 to 35%,and is associated with rice cooking and eating *** rice has low AC,generally between 6% and 15%,and its eating quality is high whether it is consumed hot or ***,the appearance quality of current soft rice cultivars needs to be improved,especially opaque *** genetic engineering has improved some agronomic traits of soft rice varieties,but not endosperm *** the present study,a RNAi construct of the soluble starch synthase Ⅱ-2(SSSⅡ-2) and the hygromycin phosphotransferase(HPT) gene were introduced into an elite japonica rice variety,Kangtiaowuyunjing(KWY8) by *** selectable marker-free(SMF) transgenic lines were obtained,and SSSⅡ-2 expression was significantly downregulated in selected transgenic lines,resulting in lower AC of the *** physicochemical properties of the transgenic rice kernels,including gel consistency(GC) and rapid visco analyzer(RVA) profile,differed significantly from those of wild-type rice and were similar to those of a soft rice variety,Nanjing 46(NJ46).These findings indicate that the cooking,eating,and processing qualities of transgenic rice are comparable to those of ***,the transgenic rice endosperm retained a transparent appearance under low-moisture ***,SMF SSSⅡ-2 RNAi rice provides a resource for breeding soft rice with transparent endosperm.

关键词： Rice(Oryza sativa L.) SSSⅡ-2 RNAi Starch quality SMF transgenic lines

RLS3, a protein with AAA+ domain localized in chloroplast, sustains leaf longevity in rice

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Journal of Integrative plant Biology 2016年第12期58卷 971-982页

作者： Yanhui Lin Lubin Tan Lei Zhao Xianyou Sun Chuanqing Sun State Key laboratory of plant Physiology and Biochemistry National Center for Evaluation of Agricultural Wild Plants (Rice) MOE Key Laboratory of Crop Heterosis and Utilization Department of Plant Genetics and Breeding China Agricultural University

Leaf senescence plays an important role in crop developmental processes that dramatically affect crop yield and grain quality. The genetic regulation of leaf senescence is complex, involving many metabolic and signaling pathways. Here, we identified a rapid leaf senescence 3 （rls3） mutant that displayed accelerated leaf senescence, shorter plant height and panicle length, and lower seed set rate than the wild type. Map-based cloning revealed that RLS3 encodes a protein with AAA＋ domain, localizing it to chloroplasts. Sequence analysis found that the rls3 8ene had a single-nucleotide substitution （G--~A） at the splice site of the Ioth intron/11th exon, resulting in the cleavage of the first nucleotide in 11th exon and premature termination of P, LS3 protein translation. Using transmission electron microscope, the chloroplasts of the rls3 mutant were observed to degrade much faster than those of the wild type. The investigation of the leaf senescence process under dark incubation conditions further revealed that the rls3 mutant displayed rapid leaf senescence. Thus, the RLS3 gene plays key roles in sustaining the normal growth of rice, while loss of function in RLS3 leads to rapid leaf senescence. The identification of RLS3 will be helpful to elucidate the mechanisms involved in leaf senescence in rice.

关键词： AAA+ domain rapid leaf senescence rice RLS3

Genome-wide Transcription Factor Gene Prediction and their Expressional Tissue-Specificities in Maize

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Journal of Integrative plant Biology 2012年第9期54卷 616-630页

作者： Yi Jiang Biao Zeng Hainan Zhao Mei Zhang Shaojun Xie Jinsheng Lai State Key laboratory of Agrobiotechnology and National Maize Improvement Center Department of Plant Genetics and BreedingChina Agricultural UniversityBeijing 100193China

Transcription factors （TFs） are important regulators of gene expression. To better understand TFencoding genes in maize （Zea mays L.）, a genome-wide TF prediction was performed using the updated B73 reference genome. A total of 2 298 TF genes were identified, which can be classified into 56 families. The largest family, known as the MYB superfamily, comprises 322 MYB and MYB-related TF genes. The expression patterns of 2014 （87.64%） TF genes were examined using RNA-seq data, which resulted in the identification of a subset of TFs that are specifically expressed in particular tissues （including root, shoot, leaf, ear, tassel and kernel）. Similarly, 98 kernel-specific TF genes were further analyzed, and it was observed that 29 of the kernel-specific genes were preferentially expressed in the early kernel developmental stage, while 69 of the genes were expressed in the late kernel developmental stage. Identification of these TFs, particularly the tissue-specific ones, provides important information for the understanding of development and transcriptional regulation of maize.

关键词： Genome-wide maize tissue-specific transcription factor.

Engineering purple rice for human health

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Science China(Life Sciences) 2018年第3期61卷 365-367页

作者： Xin Fang Yingbo Mao Xiaoya Chen National Key laboratory of plant molecular genetics CAS Center for Excellence in Molecular Plant Sciences Institute of Plant Physiology and Ecology SIBS Chinese Academy of Sciences plant Science Research Center Shanghai Key Laboratory of Plant Functional Genomics and ResourcesShanghai Chenshan Botanical Garden

Human history of domestication of wild plants as food source has witnessed continuous improvement in nutritional qualities of plant products. However, some nutritional traits were lost during agriculatral breeding （Tieman et al., 2017）, and some are absent in certain wild ancestors and could not be introduced into modem crops by traditional breeding. Micronutrient and phytonutrient deficiencies have devastat- ing effects to human health, especially to poor populations （FAO, 2013）, biofortification of food crops by molecular breeding is important to resolve these problems. Currently, there are two main strategies for biofortification, one is to increase level of already existing health-promotion com- pounds in plants, such as anthocyanin-enriched ＂Purple Tomatoes＂（Butelli et al., 2008） and folate-enriched rice （Blancquaert et al., 2015）. The other is to transfer a biosyn- thetic pathway of bioactive compounds into a plant organ absent of them, such as 13-carotene-enriched ＂Golden Rice＂（Ye et al., 2000; Paine et al., 2005）; this strategy is mostly a difficult task using conventional cloning vector systems or current transgene-stacking vector systems that enable assembly of limited numbers of target genes into single vectors with sophisticated strategies, such as using homing endonuclease sites or Multisite Gateway. Thus,

关键词：健康米饭紫色植物产品工程食物来源调制解调器繁殖期

Genetic dissection of ear-related traits using immortalized F2 population in maize

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Journal of Integrative Agriculture 2022年第9期21卷 2492-2507页

作者： GAO Ri-xin HU Ming-jian ZHAO Hai-ming LAI Jin-sheng SONG Wei-bin State Key laboratory of plant Physiology and Biochemistry&National Maize Improvement Center Department of Plant Genetics and BreedingChina Agricultural UniversityBeijing 100193P.R.China Center for Crop Functional Genomics and molecular breeding China Agricultural UniversityBeijing 100193P.R.China

Ear-related traits are often selection targets for maize improvement. This study used an immortalized F(IF) population to elucidate the genetic basis of ear-related traits. Twelve ear-related traits(namely, row number(RN), kernel number per row(KNPR), ear length(EL), ear diameter(ED), ten-kernel thickness(TKT), ear weight(EW), cob diameter(CD),kernel length(KL), kernel width(KW), grain weight per ear(GW), 100-kernel weight(HKW), and grain yield per plot(GY)),were collected from the IFpopulation. The ear-related traits were comprised of 265 crosses derived from 516 individuals of the recombinant inbred lines(RILs) under two separated environments in 2017 and 2018, respectively. Quantitative trait loci(QTLs) analyses identified 165 ear traits related QTLs, which explained phenotypic variation ranging from 0.1 to 12.66%. Among the 165 QTLs, 19 underlying nine ear-related traits(CD, ED, GY, RN, TKT, HKW, KL, GW, and KNPR)were identified across multiple environments and recognized as reliable QTLs. Furthermore, 44.85% of the total QTLs showed an overdominance effect, and 12.72% showed a dominance effect. Additionally, we found 35 genomic regions exhibiting pleiotropic effects across the whole maize genome, and 17 heterotic loci(HLs) for RN, EL, ED and EW were identified. The results provide insights into genetic components of ear-related traits and enhance the understanding of the genetic basis of heterosis in maize.

关键词： maize(Zea mays L.) QTL mapping genotyping by sequencing immortalized F2 population ear-related traits

Combining QTL mapping and expression profile analysis to identify candidate genes of cold tolerance from Dongxiang common wild rice(Oryza rufipogon Griff.)

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Journal of Integrative Agriculture 2016年第9期15卷 1933-1943页

作者： ZHAO Jie QIN Jing-jing SONG Qian SUN Chuan-qing LIU Feng-xia State Key laboratory of plant Physiology and Biochemistry National Center for Evaluation of Agricultural Wild Plants (Rice)/Beijing Key Laboratory of Crop Genetic Improvement/Department of Plant Genetics and Breeding China Agricultural University

Rice（Oryza sativa L.）, a tropical and subtropical crop, is susceptible to low temperature stress during seedling, booting, and flowering stages, which leads to lower grain quality levels and decreasing rice yields. Cold tolerance is affected by multiple genetic factors in rice, and the complex genetic mechanisms associated with chilling stress tolerance remain unclear. Here, we detected seven quantitative trait loci（QTLs） for cold tolerance at booting stage and identified one cold tolerant line, SIL157, in an introgression line population derived from a cross between the indica variety Guichao 2, as the recipient, and Dongxiang common wild rice, as the donor. When compared with Guichao 2, SIL157 showed a stronger cold tolerance during different growth stages. Through an integrated strategy that combined QTL-mapping with expression profile analysis, six candidate genes, which were up-regulated under chilling stress at the seedling and booting developmental stages, were studied. The results may help in understanding cold tolerance mechanisms and in using beneficial alleles from wild rice to improve the cold tolerance of rice cultivars through molecular marker-assisted selection.

关键词： common wild rice introgression line low temperature tolerance different growth stages candidate genes

Comparative study of adsorptive role of carbonaceous materials in removal of UV-active impurities of paclitaxel extracts

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Journal of Pharmaceutical Analysis 2015年第6期5卷 396-399页

作者： Jaber Nasiri Elaheh Motamedi Mohammad Reza Naghavi Department of Agronomy and plant breeding Division of Molecular Plant Genetics College of Agricultural & Natural Resources University of Tehran

Graphite oxide （GO） and reduced graphene oxide （rGO） nanosheets were synthesized with a low-cost manufacturing method. The morphology and structures of the synthesized samples were studied using X-ray diffraction （XRD）, atomic force microscopy （AFM）, Fourier-transform infrared （FTIR） and Raman spectroscopy. The efficiencies of GO and rGO as novel candidate adsorbents in the pre-purification of paclitaxel were compared and contrasted with those of commercial graphite （Gt）, graphene （G） and multi-wall carbon nanotube （MWCNT）. According to UV-vis and HPLC analyses, rGO was evaluated as the best absorbent for the removal of impurities in pre-purification of paclitaxel from plant cell cultures. In contrast, the GO had the poorest proficiency for paclitaxel pre-purification in comparison with the other carbonaceous adsorbents. This is attributed to the existence of many localized defects in the π-π structure of GO that is related to weakness of π-π stacking interactions between crude extract impurities and GO.

关键词： PaclitaxelReduced graphene oxideGraphite oxidePurificationCarbon nanotube

OsHAL3, a Blue Light-Responsive Protein, Interacts with the Floral Regulator Hdl to Activate Flowering in Rice

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molecular plant 2016年第2期9卷 233-244页

作者： Lei Su Jun-Xiang Shan Ji-Ping Gao Hong-Xuan Lin National Key laboratory of plant molecular genetics CAS Centre for Excellence in Molecular Plant Sciences and Collaborative Innovation Center of Genetics and Development Shanghai Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 300 Fenglin Road Shanghai 200032 China

In flowering plants, photoperiodic flowering is controlled by a complicated network. Light is one of the most important environmental stimuli that control the timing of the transition from vegetative growth to reproductive development. Several photoreceptors, including PHYA, PHYB, CRY2, and FKF1 in Arabi- dopsis and their homologs （OsPHYA, OsPHYB, OsPHYC, and OsCRY2） in rice, have been identified to be related to flowering. Our previous study suggests that OsHAL3, a flavin mononucleotide-binding protein, may function as a blue-light sensor. Here, we report the identification of OsHAL3 as a positive regulator of flowering in rice. OsHAL3 overexpression lines exhibited an early flowering phenotype, whereas down- regulation of OsHAL3 expression by RNA interference delayed flowering under an inductive photoperiod （short-day conditions）. The change in flowering time was not accompanied by altered I-Idl expression but rather by reduced accumulation of Hd3a and MADS14 transcripts. OsHAL3 and Hdl colocalized in the nucleus and physically interacted in vivo under the dark, whereas their interaction was inhibited by white or blue light. Moreover, OsHAL3 directly bound to the promoter of Hd3a, especially before dawn. We conclude that OsHAL3, a novel light-responsive protein, plays an essential role in photoperiodic control of flowering time in rice, which is probably mediated by forming a complex with Hdl. Our findings open up new perspectives on the photoperiodic flowering pathway.

关键词： Oryza sativa photoperiodic flowering blue-light-responsive protein OsHAL3 Hdl

Histone 3 lysine 36 to methionine mutations stably interact with and sequester SDG8 in Arabidopsis thaliana

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Science China(Life Sciences) 2018年第2期61卷 225-234页

作者： Guang Lin Ying Zhou Min Li Yuda Fang National key laboratory of plant molecular genetics Chinese Academy of Sciences Center for Excellence in Molecular Plant SciencesInstitute of Plant Physiology and Ecology Chinese Academy of Sciences University of Chinese Academy of Sciences Shanghai 200032 China

Post-transcriptional modifications of histones play important roles in various biological processes. Here, we report that Arabidopsis plants overexpressing histone H3 lysine to methionine mutations at histone H3.1K36(H3.1K36M) and H3.3K36(H3.3K36M) have serious developmental defects with early-flowering and change in the modifications of endogenous histone H3, including acetylation at lysine 9(H3K9ac), trimethylation at lysine 27(H3K27me3), di-and tri-methylation at lysine 36(H3K36me2 and H3K36me3). In addition, H3K36M mutation alters its subcellular localization and interacts with H3K36 methyltransferase SDG8. Our results support a model in which H3K36M stably interacts with SDG8, and inhibits the activity of SDG8 by sequestering SDG8, resulting in a dominant negative effect to affect the proper expression levels of a variety of genes and plant development.

关键词： histone H3 lysine to methionine mutation SDG8 dominant negative effect Arabidopsis