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1. 黄瓜顶端分生和花芽组织的碳蜡原位杂交体系的优化
WANG Cui, SUN Jin-jing, YANG Xue-yong, WAN Li, ZHANG Zhong-hua, ZHANG Hui-min
Journal of Integrative Agriculture    2023, 22 (2): 464-470.   DOI: 10.1016/j.jia.2022.08.038
摘要176)      PDF    收藏

黄瓜是研究花发育的重要模式作物,控制花发育相关的基因通常具有特异的时空表达模式,原位杂交技术(ISH)被广泛应用在分析黄瓜茎端分生组织(SAM)和花芽组织FM的基因时空表达模式上。原位杂交的常用方法是石蜡原位杂交,然而石蜡原位杂交中组织包埋周期长,RNA容易降解,导致信号减弱。本研究基于石蜡原位杂交进行了优化建立了碳蜡原位杂交方法,具有操作简便,灵敏度高的优点。该方法一是使用低熔点的碳蜡,RNA保存好;二是结合了整体原位杂交的方法,在溶液中处理组织切片,解决了碳蜡不易粘片的问题。在文章中,利用碳蜡原位杂交方法,我们在黄瓜的茎端分生组织(SAM)和花芽组织FM)中检测了CLV3WUS1基因的表达模式,实验结果与前期研究结果一致,而且该方法比石蜡原位杂交周期缩短4-5天,信号强。同样地,该方法也成功用于拟南芥中CLV3WUS1的表达模式分析表明碳蜡原位杂交方法在大多数物种中具有普遍适用性。

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2. ACS11一个新突变类型导致黄瓜只开雄花
WANG Jie, LI Shuai, CHEN Chen, ZHANG Qi-qi, ZHANG Hui-min, CUI Qing-zhi, CAI Guang-hua, ZHANG Xiao-peng, CHAI Sen, WAN Li, YANG Xue-yong, ZHANG Zhong-hua, HUANG San-wen, CHEN Hui-ming, SUN Jin-jing
Journal of Integrative Agriculture    2023, 22 (11): 3312-3320.   DOI: 10.1016/j.jia.2023.03.003
摘要199)      PDF    收藏

植物的单性花可以有效促进异交,研究单性花的形成和调控机制对于理解植物性别决定过程有重要意义,也为研究者和农业生产者利用杂种优势提供便利。在黄瓜杂交制种过程中,只开花的株系种植于只开雄花的株系周围,可以显著降低制种成本。筛选更多不同基因背景的只开雄花的材料,将增加可用于育种的种质资源。我基于前期构建的EMS诱变自交系材料406”的突变体库,发现了一个新的只开雄花的突变体遗传分析、全基因组重测序和分子标记辅助验证表明,ACS11基因上发生异义突变301位丝氨酸(Ser)变为苯丙氨酸(Phe)导致全雄株的产生。体外酶活性测定表明,此突变导致酶活性完全丧失。本研究为黄瓜雄性亲本选育提供了新的种质资源,并为 ACS 酶的催化机理提供了新的认识。

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3.
Allelochemical p-hydroxybenzoic acid inhibits root growth via regulating ROS accumulation in cucumber (Cucumis sativus L.)
HUANG Cheng-zhen, XU Lei, Sun Jin-jing, ZHANG Zhong-hua, FU Mei-lan, TENG Hui-ying, YI Ke-ke
Journal of Integrative Agriculture    2020, 19 (2): 518-527.   DOI: 10.1016/S2095-3119(19)62781-4
摘要90)      PDF    收藏
Allelopathy is prevalent in agricultural ecosystems and mediated by plant-derived secondary metabolites (allelochemicals).  Allelochemicals are released by donor plants and affect the root growth and development of receptor plants.  Allelopathy is responsible for the continuous cropping obstacles in cucumber (Cucumis sativus L.).  p-Hydroxybenzoic acid (pHBA), an autotoxin from root exudates of cucumber, has been proposed to be an important allelopathic chemical.  However, the molecular mechanism by which pHBA affect root growth and development in cucumber is unknown.  Here, we found that pHBA treatment suppressed root growth of cucumber by reducing the meristem activity and cell length.  This root growth defect is caused by reduced reactive oxygen species (ROS) accumulation in root tips.  After pHBA treatment, the expression levels of several ROS-scavenging-related genes were increased, including peroxidase (POD), catalase (CAT) and metallothionein (MT).  Moreover, exogenously application of salicylhydroxamate (SHAM), a peroxidase inhibitor, can partially restore the pHBA treatment induced root growth inhibition.  Furthermore, we found that there is natural variation for the inhibitory effect of pHBA on root growth.  We also showed that pHBA treatment could maintain higher level of ROS accumulated in the pHBA less sensitive cucumber than that in the pHBA-sensitive cucumber.  These results suggest that pHBA inhibits root growth by reducing root tip ROS level in cucumber.
 
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4. Analysis of SSRs Information in Capsicum spp. from EST Database
HUANG Huan-huan, ZHANG Zhong-hua, ZHANG Zheng-hai, MAO Sheng-li, WANG Li-hao , ZHANG Bao-xi
Journal of Integrative Agriculture    2011, 10 (10): 1532-1536.   DOI: 10.1016/S1671-2927(11)60148-X
摘要1988)      PDF    收藏
SSR markers are useful in pepper linkage mapping and gene location. 446 SSR markers have been reported, but they are insufficient. It is costly to develop SSR markers from DNA library, whereas it seems much easy to find in EST sequences in the GenBank of pepper through internet. In this study, attempts have been made to develop SSR markers in the EST sequences by using bioinformatics. EST sequences were trimmed by ‘est-trimmer.pl’ software, while 116915 EST sequences were obtained without poly ‘A’ or poly ‘T’, ranged between 100 and 700 bp. Using ‘e-PCR’ and ‘del.pl’ softwares, SSR sequences were identified. 2 508 microsatellite loci (larger than 20 repeats) were established and 755 SSR primers were designed using SSR finder software and Primer 3 software. There were 498 (0.43%) mono-, 1 026 (0.89%) di-, 518 (0.45%) tri-, 245 (0.21%) tetra-, 114 (0.10%) penta-, and 107 (0.09%) hexa-nucleotide SSRs. The estimated frequency of SSRs was approximately 1/25.12 kb. According to the distribution of SSRs in pepper, the mean length of pepper SSRs was 22.68 bp and the adenine rich repeats such as A/T, AG, AT, AAG, AAAT, and AAAC were predominant in each type of SSRs (mono-, di-, tri-, tetra-, penta-, and hexa-), whereas the C/G, CG, CCG repeats were less abundant. 210 primers were tested in 8 pepper cultivars and the PCR result revealed the existence of polymorphism among 127 (60.48%) SSR primers within 8 pepper cultivars. It is confirmed that pepper EST database could be efficiently exploited for availability of SSR markers.
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