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1. JIA-2021-1609 基于耗氧量和顶空气相色谱-离子迁移谱技术的单粒种子生活力判定
TU Ke-ling, YIN Yu-lin, YANG Li-ming, WANG Jian-hua, SUN Qun
Journal of Integrative Agriculture    2023, 22 (3): 727-737.   DOI: 10.1016/j.jia.2022.08.058
摘要301)      PDF    收藏
鉴定和筛选优质种子对于提高作物产量至关重要。本研究的目的是通过预测单粒种子的潜在发芽能力,判定种子是否具有生活力,以改进作物种子的精选技术,从而提高种子质量。本研究基于种子萌发早期的耗氧量(Q)和种子的挥发性气体成分对单粒种子的生活力进行判定。甜玉米种子、辣椒、小麦种子,有生活力和无生活力种子,其呼吸特征值——最终耗氧量(Q120)均存在差异。在此基础上,为了缩短Q2技术的测量过程,尽可能地减少测量过程中对种子造成不可逆的损害,我们探究了新的耗氧量变量Qt,将辣椒、甜玉米和小麦种子生活力评估的时间分别提前到12、6和9小时,生活力预测的准确率分别为91.9%、97.7%和96.2%。此过程可摒弃无生活力的种子,从而提高种子批的质量,将三种作物的发芽率分别从原始的86.6%、90.9%和53.8%提高到100%。考虑到单粒甜玉米种子的质量较大,存在从单粒种子上检测到挥发性气体成分的可能性,我们进一步尝试了将空气相色谱-离子迁移谱技术(HS-GC-IMS)应用于单粒甜玉米种子的生活力判定,共鉴定出48个峰,其中38种挥发性物质被表征,包括醇、醛、酸和酯。然而,由于单粒甜玉米种子之间挥发性气体成分的差异极为细微,有生活力种子和无生活力种子之间未检测到显著差异,基于挥发性气体成分的主成分分析也无法区分这两种类型的甜玉米种子。本研究所采取的这些方法可为单粒种子生活力的快速鉴定提供参考。
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2. 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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3. Effect of mechanical threshing on damage and vigor of maize seed threshed at different moisture contents
GU Ri-liang, HUANG Ran, JIA Guang-yao, YUAN Zhi-peng, REN Li-sha, LI Li, WANG Jian-hua
Journal of Integrative Agriculture    2019, 18 (7): 1571-1578.   DOI: 10.1016/S2095-3119(18)62026-X
摘要272)      PDF    收藏
Mechanical threshing used when preparing maize seeds for planting subjects seed to damage and reduces seed quality.  The purpose of this study was to assess the effect of mechanical threshing on the quality of maize seed threshed at different moisture contents (MCs).  Seeds of dent maize JK968 and flint maize DD2 were threshed at 12, 15, 18, 21, and 24% MC.  The damage degree was determined by iodine staining, and seed vigor was assessed by standard germination (SG), cold test germination (CTG), accelerated aging germination (AAG), seedling emergence rate, and seedling root and shoot length.  The results showed that the damage percentage increased, and the seed vigor parameters decreased with increasing seed MC during threshing in both cultivars.  For obtaining high seed quality, indicated by at least 90, 85, and 80% of SG, AAG, and CTG, respectively, JK 968 and DD2 should be threshed at MC lower than 15 and 18%, respectively.  Furthermore, the damage mainly occurred in the apical part of seeds, irrespective of the threshing MC in both cultivars.  When the embryo was damaged, seedling emergence rates were significantly reduced with great influence on shoot length.  Damage to the endosperm resulted in little effect on seedling performance.  Flint maize DD2 was more tolerant to mechanical threshing than dent maize JK968.  These results provided technical reference for the production and processing of high vigor maize seeds.
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4. Selection for high quality pepper seeds by machine vision and classifiers
TU Ke-ling, LI Lin-juan, YANG Li-ming, WANG Jian-hua, SUN Qun
Journal of Integrative Agriculture    2018, 17 (09): 1999-2006.   DOI: 10.1016/S2095-3119(18)62031-3
摘要425)      PDF    收藏
This research aimed to improve selection of pepper seeds for separating high-quality seeds from low-quality seeds. Past research has shown that seed vigor is significantly related to the seed color and size, thus several physical features were identified as candidate predictors of high seed quality. Image recognition software was used to automate recognition of seed feature quality using 400 kernels of pepper cultivar 101. In addition, binary logistic regression and a neural network were applied to determine models with high predictive value of seed germination. Single-kernel germination tests were conducted to validate the predictive value of the identified features. The best predictors of seed vigor were determined by the highest correlation observed between the physical features and the subsequent fresh weight of seedlings that germinated from the 400 seeds. Correlation analysis showed that fresh weight was significantly positively correlated with eight physical features: three color features (R, a*, brightness), width, length, projected area, and single-kernel density, and weight. In contrast, fresh weight significantly negatively correlated with the feature of hue. In analyses of two of the highest correlating single features, germination percentage increased from 59.3 to 71.8% when a*≥3, and selection rate peaked at 57.8%. Germination percentage increased from 59.3 to 79.4%, and the selection rate reached 76.8%, when single-kernel weight ≥0.0064 g. The most effective model was based on a multilayer perceptron (MLP) neural network, consisting of 15 physical traits as variables, and a stability calculated as 99.4%. Germination percentage in a calibration set of seeds was 79.1% and the selection rate was 90.0%. These results indicated that the model was effective in predicting seed germination based on physical features and could be used as a guide for quality control in seed selection. Automated systems based on machine vision and model classifiers can contribute to reducing the costs and labor required in the selection of pepper seeds.
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5. Overexpression of a maize SNF-related protein kinase gene, ZmSnRK2.11, reduces salt and drought tolerance in Arabidopsis
ZHANG Fan, CHEN Xun-ji, WANG Jian-hua, ZHENG Jun
Journal of Integrative Agriculture    2015, 14 (7): 1229-1241.   DOI: 10.1016/S2095-3119(14)60872-8
摘要1976)      收藏
Sucrose non-fermenting-1 related protein kinase 2 (SnRK2) is a unique family of protein kinases associated with abiotic stress signal transduction in plants. In this study, a maize SnRK2 gene ZmSnRK2.11 was cloned and characterized. The results showed that ZmSnRK2.11 is up-regulated by high-salinity and dehydration treatment, and it is expressed mainly in maize mature leaf. A transient expression assay using onion epidermal cells revealed that ZmSnRK2.11-GFP fusion proteins are localized to both the nucleus and cytoplasm. Overexpressing-ZmSnRK2.11 in Arabidopsis resulted in salt and drought sensitivity phenotypes that exhibited an increased rate of water loss, reduced relative water content, delayed stoma closure, accumulated less free proline content and increased malondialdehyde (MDA) content relative to the phenotypes observed in wild-type (WT) control. Furthermore, overexpression of ZmSnRK2.11 up-regulated the expression of the genes ABI1 and ABI2 and decreased the expression of DREB2A and P5CS1. Taken together, our results suggest that ZmSnRK2.11 is a possible negative regulator involved in the salt and drought stress signal transduction pathways in plants.
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6. Functional Analysis of the ZAG2 Promoter from Maize in Transgenic Tobaccos
LU Min-hui, WANG Guo-ying, MENG Zheng, WANG Jian-hua
Journal of Integrative Agriculture    2012, 12 (8): 1266-1273.   DOI: 10.1016/S1671-2927(00)8655
摘要1351)      PDF    收藏
The function of the 3 040 bp sequence at the upstream translation starting site (ATG) of the ZAG2 gene, isolated from the maize genome, was studied. The sequence analysis showed that the sequence contained a typical class C MADS-box gene regulatory element. The 5´ UTR region of the gene contains a 1 299-bp intron that might have important regulatory functions. To study the sequence function, deletion derivatives of promoter-reporter (uidA) gene fusions were generated and transformed into tobaccos. The GUS staining and fluorescence quantification results showed that the GUS activity was detected only in the third and fourth whorl floral organs of the transgenic tobaccos under driving the promoter including the first intron, while detected in all the organs and was stronger under driving the promoter without the first intron. However, the GUS activity was just detected in one whorl of the fourth or third floral organs under driving of the 35S promoter. These results suggested that the first intron of the ZAG2 gene contains functional regulatory elements, which turned out to be important for gene expression in the heterologous systems. Moreover, the GUS activity was decreased when the reporter gene driven by the promoters with 5´-deletions, respectively, from -1 606 to -951 and -951 to -426 nts, which indicates that positive regulatory elements are present in these two sequence stretches.
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