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1. Dissecting the genetic basis of maize deep-sowing tolerance by combining association mapping and gene expression analysis
YANG Yue, MA Yu-ting, LIU Yang-yang, Demar LYLE, LI Dong-dong, WANG Ping-xi, XU Jia-liang, ZHEN Si-han, LU Jia-wen, PENG Yun-ling, CUI Yu, FU Jun-jie, DU Wan-li, ZHANG Hong-wei, WANG Jian-hua
Journal of Integrative Agriculture    2022, 21 (5): 1266-1277.   DOI: 10.1016/S2095-3119(21)63649-3
摘要150)      PDF    收藏

深播是包括玉米在内的作物躲避干旱的一种重要策略,候选基因的克隆是进行玉米耐深播分子机理研究的基础工作。本研究中,我们利用包含386份玉米自交系的关联分析群体对10厘米和20厘米播种深度条件下的四个性状进行鉴定。利用50万SNP标记进行关联分析发现了273个耐深播性状显著关联的SNP。对两组不同的处理进行RNA测序分析分别发现1944和2098个差异基因,其中包含281个共同的差异基因。通过比较273个SNP和281个差异基因的位置发现了7个可能与耐深播相关的候选基因,其中GRMZM2G119769编码一个SNF1激酶互作的蛋白。由于GRMZM2G119769在其他植物中的同源基因跟器官伸长、生长素和光响应有关。同时,候选基因关联分析表明GRMZM2G119769基因的自然变异与玉米的中胚轴长度有关。另外,基因表达分析表明GRMZM2G119769在耐深播材料中表达量高。这些研究结果都支持GRMZM2G119769是玉米耐深播性状的候选基因。本研究不但评价了玉米资源的耐深播特性,还鉴定出一些可能对未来玉米耐深播研究有参考价值的候选基因。


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2. Breeding of CMS maintainer lines through anther culture assisted by high-resolution melting-based markers
WANG Ping, BAI Yu-lu, WANG Min-xia, HU Bin-hua, PU Zhi-gang, ZHANG Zhi-yong, ZHANG Qiong, XU Deng-wu, LUO Wen-long, CHEN Zhi-qiang
Journal of Integrative Agriculture    2020, 19 (12): 2965-2973.   DOI: 10.1016/S2095-3119(20)63179-3
摘要101)      PDF    收藏
The integrated use of molecular marker-assisted selection (MAS) and anther culture has potential to significantly increase efficiency in plant breeding; however, reports on this kind of practical use are very limited.  In the present study, we report the development of cytoplasmic male sterile (CMS) maintainers with aroma, disease resistance and red-brown hulls, as an example of integration of MAS and anther culture in rice breeding.  A high-resolution melting (HRM)-based functional molecular marker was developed for the red-brown hull trait caused by a unique mutation (rbh1) in OsCAD2.  Functional molecular markers for genes of rice blast resistance (Pi2), aroma (fgr) and red-brown hull (rbh1) were used for precise genotyping of individual plants in the BC1 and BC2F2 populations derived from a cross between CMS maintainers Huaxiang B (pi2–/rbh1–/fgr–) and Rong 3B (Pi2+/RBH1+/Fgr+).  A total of 89 doubled haploid (DH) lines were generated from selected BC2F2 plants (Pi2+/rbh1–/fgr–) by anther culture.  Seven DH lines were subsequently selected as the potential new CMS maintainers based on their overall performance and high resistance to blast.  Our study demonstrated that integration of MAS and anther culture significantly accelerated the development of CMS maintainers with multiple stacked genes.
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3. Genome-wide association study for starch content and constitution in sorghum (Sorghum bicolor (L.) Moench)
CHEN Bing-ru, WANG Chun-yu, WANG Ping, ZHU Zhen-xing, XU Ning, SHI Gui-shan, YU Miao, WANG Nai, LI Ji-hong, HOU Jia-ming, LI Shu-jie, ZHOU Yu-fei, GAO Shi-jie, LU Xiao-chun, HUANG Rui
Journal of Integrative Agriculture    2019, 18 (11): 2446-2456.   DOI: 10.1016/S2095-3119(19)62631-6
摘要124)      PDF    收藏
Starch is the most important component in endosperm of sorghum grain.  Usually, two types of starch are present: amylose (AM) and amylopectin (AP).  The levels of AM and AP contents play a significant role in the appearance, structure, and quality of sorghum grains and in marketing applications.  In the present study, a panel of 634 sorghum (Sorghum bicolor (L.) Moench) accessions were evaluated for starch, AM, and AP contents of grain, which included a mini core collection of 242 accessions from the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) in India, and 252 landraces and 140 cultivars from China.  The average starch content was 67.64% and the average AM and AP contents were 20.19 and 79.81%, respectively.  We developed a total of 260 000 high-confidence single nucleotide polymorphism (SNP) markers in the panel of 634 accessions of S. bicolor using specific locus amplified fragment sequencing (SLAF-seq).  We performed genome-wide association studies (GWAS) of starch, AM, and AM/AP of grain and SNP markers based on a mixed linear model (MLM).  In total, 70 significant association signals were detected for starch, AM, and AM/AP ratio of grain with P<4.452×10–7, of which 10 SNPs were identified with significant starch, 51 SNPs were associated with AM, and nine SNPs were associated with the AM/AP ratio.  The Gene Ontology (GO) analysis identified 12 candidate genes at five QTLs associated with starch metabolism within the 200-kb intervals, located on chromosomes 1, 5, 6, and 9.  Of these genes, Sobic.006G036500.1 encodes peptidyl-prolyl cis-trans-isomerase CYP38 responsible for hexose monophosphate shunt (HMS) and Sobic.009G071800 encodes 6-phospho-fructokinase (PFK), which is involved in the embden-meyerhof pathway (EMP).  Kompetitive allele specific PCR (KASP) markers were developed to validate the GWAS results.  The C allele is correlated with a high starch content, while the T allele is linked with a low level of starch content, and provides reliable haplotypes for MAS in sorghum quality improvement.
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4. Analysis of the diversity and function of the alleles of the rice blast resistance genes Piz-t, Pita and Pik in 24 rice cultivars
WANG Yan, ZHAO Jia-ming, ZHANG Li-xia, WANG Ping, WANG Shi-wei, WANG Hui, WANG Xiao-xi, LIU Zhi-heng, ZHENG Wen-jing
Journal of Integrative Agriculture    2016, 15 (7): 1423-1431.   DOI: 10.1016/S2095-3119(15)61207-2
摘要1271)      PDF    收藏
Understanding the sequence diversity of rice blast resistance genes is important for breeding new resistant rice cultivars against the rice blast fungus Magnaporthe oryzae. In this study, we selected 24 rice cultivars with different genetic backgrounds to study the allelic diversity of rice blast resistance genes Piz-t, Pita and Pik. For Piz-t, a total of 17 allelic types were found within the 24 cultivars. Blast inoculations showed that most of the mutations can affect the function of the resistance gene. For Pita, except for the difference at the 918th amino acid, a majority of the 21 mutations were detected among the cultivars. Inoculations with blast isolates carrying Avr-Pita revealed that cultivars with mutations in other sites except for the 918th amino acid did not affect the function of the Pita gene. For Pik, a total of six allelic types were found within the 24 cultivars, but five of them lost the function of the resistance gene. In addition, we found that Piz-t, Pita and Pik were expressed constitutively in the 24 rice cultivars and the expression level was not related to resistance. Our results have provided the sequence diversity information of the resistance genes Piz-t, Pita and Pik among the popular rice cultivars grown in the northeast region of China.
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5. Foliar application of sodium hydrosulfide (NaHS), a hydrogen sulfide (H2S) donor, can protect seedlings against heat stress in wheat (Triticum aestivum L.)
YANG Min, QIN Bao-ping, MA Xue-li, WANG Ping, LI Mei-ling, CHEN Lu-lu, CHEN Lei-tai, SUN Aiqing, WANG Zhen-lin, YIN Yan-ping
Journal of Integrative Agriculture    2016, 15 (12): 2745-2758.   DOI: 10.1016/S2095-3119(16)61358-8
摘要1312)      PDF    收藏
    Temperature extremes represent an important limiting factor to plant growth and productivity. Low concentration of hydrogen sulfide (H2S) has been proven to function in physiological responses to various stresses. The present study evaluated the effect of foliar application of wheat seedlings with a H2S donor, sodium hydrosulfide (NaHS), on the response to acute heat stress. The results showed that pretreatment with NaHS could promote heat tolerance of wheat seedlings in a dose-dependent manner. Again, it was verified that H2S, rather than other sulfur-containing components or sodion derived from NaHS solution, should contribute to the positive role in promoting wheat seedlings against heat stress. To further study antioxidant mechanisms of NaHS-induced heat tolerance, superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6) and ascorbate peroxidase (APX, EC 1.11.1.11) activities, and H2S, hydrogen peroxide (H2O2), malonaldehyde (MDA), and soluble sugar contents in wheat seedlings were determined. The results showed that, under heat stress, the activities of SOD, CAT, and APX, H2S, H2O2, MDA, and soluble sugar contents in NaHS-pretreated seedlings and its control all increased. Meanwhile, NaHS-pretreated seedlings showed higher antioxidant enzymes activities and gene expression levels as well as the H2S and soluble sugar levels, and lower H2O2, MDA contents induced by heat stress. While little effect was detected in antioxidant enzymes activities and soluble substances contents in pretreated wheat seedlings compared with its control under normal culture conditions (data not shown). All of our results suggested that exogenous NaHS could alleviate oxidative damage and improve heat tolerance by regulating the antioxidant system in wheat seedlings under heat stress.
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6. EaIspF1, Essential Enzyme in Isoprenoid Biosynthesis from Eupatorium adenophorum, Reveals a Novel Role in Light Acclimation
ZHANG Sheng-rui, JIANG Xue, WANG Ping, WU Di, WANG Qing-hua , HOU Yu-xia
Journal of Integrative Agriculture    2014, 13 (5): 1024-1035.   DOI: 10.1016/S2095-3119(13)60519-5
摘要2131)      PDF    收藏
Isoprenoids are a functionally and structurally diverse class of natural organic chemicals. The universal precursors of all isoprenoids, isopentenyl diphosphate and dimethylallyl diphosphate are synthesized through the mevalonate and 2C-methyl- D-erythritol 4-phosphate (MEP) pathways, respectively. Many isoprenoids produced through the MEP pathway play an important role in plant acclimation to different light environments. Eupatorium adenophorum, an invasive weed in China, presents a remarkable capacity to acclimate to various light environments, which constitutes its solid foundation of being a successful invasive species. Thus we aimed at gaining a deeper insight into the regulation of MEP pathway in E. adenophorum to further understand the invasive mechanism. 2C-Methyl-D-erythritol 2,4-cyclodiphosphate synthase (IspF or MCS) is an essential enzyme in the MEP pathway. In this paper, a novel IspF gene was cloned and characterized from E. adenophorum. Tissue-specific expression assays revealed a higher expression of EaIspF1 in leaves than in stems and roots. The expression of EaIspF1 was responsive to different light conditions. Some up-regulation of EaIspF1 expression was also found after the treatments with signal compounds and after wounding stress. Interestingly, the over-expression of EaIspF1 in Arabidopsis led to increase carotenoids contents, resulting in an enhanced tolerance to high light. Taken together, these results indicate that the EaIspF1-derived enzyme participates in isoprenoid metabolism and among others, the expression of this gene in E. adenophorum is involved in the regulation of plastidial isoprenoids, which play an important role in acclimation to various light environments.
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7. Dramatic Changes of Matrix Metalloproteinases-7 and Lysozyme in the Ulcerative Colitis of Mice Induced by Dextran Sulfate Sodium
KANG Jing-jing, ZHAO De-ming, TENG Ke-dao, JIAO Xi-lan, WANG Ping-li, SUN Zhe, NI Peipei, WANG Zhi-feng, ZHANG Rui, YANG Yu-rong , LIANG Hong-de
Journal of Integrative Agriculture    2014, 13 (4): 858-859.   DOI: 10.1016/S2095-3119(13)60416-5
摘要1637)      PDF    收藏
Ulcerative colitis (UC) is a lifelong illness with profound emotional and social impacts, and could cause serious damage to large intestine, especially in colon. However, the pathogenesis of UC remained unclear. The present study attempts to find out the role of matrix metalloproteinases-7 (MMP-7) and lysozyme in the pathogenesis of UC through a mice model induced by dextran sulfate sodium (DSS). The UC model was evaluated both by disease activity index (DAI) and the intestinal histopathology. The results show that there is a high correlation between the DAI score and the pathological changes of colon. Interleukin-6 (IL-6) serum levels and large intestinal fluids levels in UC mice are always higher than that of the control groups, which might be associated with the degree of the inflammation damage in the colon. The change tendency of the MMP-7 mRNA and protein expressions are both up-regulated firstly and then down-regulated from 1 to 5 d in the colon, but only the MMP-7 protein is up-regulated at 7 d again. The up-regulated MMP-7 levels in the early stage of UC may play a protective role through the activated defensins, while the down-regulated levels in the mid-later stage of UC may be connected with the severe lesions in the colon. However, the up-regulated MMP-7 levels in the later stage of UC in the colon may also contribute to the tissue repair or be served as a marker to CRC (colorectal cancer). The distribution of lysozyme protein indicates that there may be Paneth-like cells in the colon. Both the changes of MMP-7 and lysozyme in the small intestine may play a protective role for the safe environment of the whole gut, especially to the colon of UC.
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8. Nitric Oxide Content in Wheat Leaves and Its Relation to Programmed Cell Death of Main Stem and Tillers Under Different Nitrogen Levels
GUO Jun-xiang, CHEN Er-ying, YIN Yan-ping, WANG Ping, LI Yong, CHEN Xiao-guang, WU Guanglei
Journal of Integrative Agriculture    2013, 12 (2): 239-250.   DOI: 10.1016/S2095-3119(13)60223-3
摘要1622)      PDF    收藏
Nitric oxide (NO) is a key signaling molecule in different physiological processes of plants, including programmed cell death (PCD). PCD of tillers plays an important role in surviving which are major components of grain yield. PCD was triggered in wheat leaves of main stem and tillers by NO content under different nitrogen treatments. In wheat, NO could be synthesized endogenously by nitrate reductase (NR). As an inducible enzyme, NR activity was closely related to substrate concentration. Therefore, different nitrogen levels would change NR activity and NO production. The objective of this study was to determine the effects of NR activity, NO production, and the correlation between them on different tillers growth, development, senescence, and kernel protein content under different nitrogen levels. Field-experiments were conducted in 2009-2011 growing seasons, using two wheat cultivars with different spike-types. Results showed that for main stem and primary tillers, NR activity and NO content reached high level at heading stage, while for secondary tiller, the level of NR activity was low, but NO content was high in the present research. The NO synthesis depending on NR activity in wheat leaves was significant in the early growing stage, but the NO synthesis weakened with the progress of growing period. NO was related to the senescence of wheat leaves, but PCD was more sensitive to marked changes of NO content than NO content itself. N application had marked influence on the aging process of primary tiller, while had little influence on that of main stem and secondary tiller. Moreover, N fertilizer application could increase spike rate and protein content of primary tiller by N fertilizer application.
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9. DNA Polymorphisms of 5´-Flanking Region of Insulin-Like Growth Factor 1 Gene and Their Association with Reproduction Traits in Goats 
WANG Ping-qing, TAN Ying, ZHANG Bao-yun, CHU Ming-xing, DENG La-mei, FAN Qi , LIUChong-xu
Journal of Integrative Agriculture    2011, 10 (10): 1609-1617.   DOI: 10.1016/S1671-2927(11)60158-2
摘要1848)      PDF    收藏
Research on the identity of genes and their relationship with traits of economic importance in farm animals could assist in the selection of livestock. In this study, the polymorphisms of insulin-like growth factor 1 (IGF1) gene in 561 goats of ten breeds were detected by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) and their association with litter size and birth weight in three breeds were investigated. The effects of IGF1 polymorphisms on the breeding value for litter size and birth weight were examined using least square methods. Two deletions (CA) were detected in the microsatellite and two mutations (A1637G, T1640C) were found in 5´-flanking regulatory region. No significant association between the polymorphisms in 5´-flanking region of IGF1 and birth weight was found in the three breeds of goats. In Gulin Ma goats, two polymorphisms were found to affect litter size traits. In Chuandong White goats and Guizhou White goats, no significant difference (P<0.05) in litter size between goats carrying different genotypes was observed. Further evaluation and confirmation studies in more goat populations with larger sample sizes are necessary.
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