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1. 腐烂茎线虫中药材群体ITS单倍型分析
NI Chun-hui, HAN Bian, LIU Yong-gang, Maria MUNAWAR, LIU Shi-ming, LI Wen-hao, SHI Ming-ming, LI Hui-xia, PENG De-liang
Journal of Integrative Agriculture    2023, 22 (6): 1763-1781.   DOI: 10.1016/j.jia.2022.08.126
摘要181)      PDF    收藏

腐烂茎线虫(Ditylenchus destructor)是一种重要的检疫性病原线虫,严重危害甘薯、马铃薯和中药材等根茎类作物。该线虫种内分化明显,根据ITS-rDNA序列差异,国外的研究将其分为A-G 7个单倍型,主要集中于马铃薯和甘薯的线虫群体。本研究对腐烂茎线虫中药材群体ITS-rDNA序列及其RNA二级结构进行分析以明确单倍型分化,并通过ITS-rDNA28S D2-D3系统发育关系、ITS-RFLRITS特异性引物PCR扩增进一步验证不同群体单倍型分化。甘肃、青海、陕西、内蒙和黑龙江等5个省采集当归、党参、马铃薯和甘薯的腐烂茎线虫群体43个,其中中药材群体37。线虫群体的ITS-rDNA序列长度为727 bp-969 bp,长度差异主要表现在ITS1区串联重复序列的数量不同串联重复序列在ITS1二级结构中形成了稳定的茎环,即H9螺旋。H9螺旋结构的有无及其差异,43群体的ITS-rDNA序列可划分10个单倍型与已知单倍型(A-G)对比分析,发现其中3单倍型分别AB和C单倍型一致,而另外7单倍型与已知单倍型不同,将其依次命名为HIJKLMN单倍型,7单倍来源于中药材。综合本研究和已知单倍型分体系,腐烂茎线虫中发现A-N 14个单倍型。ITS28S系统发育分析显示,所有单倍型群体分化为两支:A单倍型为一支,B-N单倍型为一支。对比ITS28S系统发育,发现A单倍型均单独聚为一支,但B-N分不太一致且不同单倍型系统发育关系较为混乱。ITS-RFLP和特异性引物PCR结果显示H和A单倍型酶切图谱和特异性片段长度相同, B和C单倍型特异性片段长度相同,但其它单倍型间存在明显差异。除K单倍型不同群体间有差异外,其单倍型群体间明显差异。本研究发现了腐烂茎线虫中药材群体中存在新的单倍型,并明确了不同单倍型差异,该结果将推动茎线虫生物学的研究进展,且对中药材腐烂茎线虫的识别和防治具有指导意义。

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2. Resistance of barley varieties to Heterodera avenae in the Qinghai–Tibet Plateau, China
Yan Jia-hui, Jia Jian-ping, JIANG Li-ling, Peng De-liang, Liu Shi-ming, Hou Sheng-ying, YU Jing-wen, Li Hui-xia, Huang Wen-kun
Journal of Integrative Agriculture    2022, 21 (5): 1401-1413.   DOI: 10.1016/S2095-3119(21)63769-3
摘要149)      PDF    收藏

禾谷孢囊线虫H. avenae是一种重要的植物病原线虫,严重影响禾谷类作物的产量。目前已在我国河南、河北、江苏、青海、西藏等16个省(市)发生危害。本研究通过人工接种试验和田间试验,利用抽穗期根系中的线虫数量指标评价了青海栽培二棱大麦 (QH2R)、青海栽培六棱大麦 (QH6R) 和西藏栽培二棱大麦 (TB2R) 对禾谷孢囊线虫的抗病性,并通过接种试验和显微观察,评价了两个高抗品种中线虫的侵染和发育情况。为更好地评价不同品种对H. avenae的抗感性,首先比较了两种常用的抗性评价方法——繁育系数法 (PPR) 和单株雌虫/孢囊数量法 (NFP) 的准确性。对田间自然条件下186个品种受害情况的调查结果表明,利用NFP法鉴定出的感病品种数量显著高于PPR法鉴定的感病品种数量,表明NFP法更利于鉴定大麦品种的抗病性。通过2015年至2017年的田间试验及2018年的人工接种试验,发现QH2R系列品种中形成的雌虫/孢囊数量最少,显著低于QH6R和TB2R系列品种。综合接种试验与田间试验的结果,从QH2R系列品种中鉴定出8个高抗品种 (Sunong7617, Sunong7635, Dongyuan87-14, Rudong14-46, Rudong87-57, Rudong87-8-45, Rudong88-14-2, Rudong88-67-1),平均单株孢囊数量低于4.2个。对线虫发育进程的显微观察表明,高抗品种 (Sunong7635和Dongyuan87-14) 中H. avenae幼虫的侵入数量显著低于感病品种中幼虫的侵入数量,并且幼虫发育成雌虫的数量也显著减少。本研究中鉴定的高抗品种对于育种工作者培育禾谷孢囊线虫抗性品种、更加经济有效地控制禾谷孢囊线虫的危害等具有重要意义


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3. High potassium to magnesium ratio affected the growth and magnesium uptake of three tomato (Solanum lycopersicum L.) cultivars
LI Hui-xia, CHEN Zhu-jun, ZHOU Ting, LIU Yan, ZHOU Jian-bin
Journal of Integrative Agriculture    2018, 17 (12): 2813-2821.   DOI: 10.1016/S2095-3119(18)61949-5
摘要362)      PDF    收藏
Potassium (K) and magnesium (Mg) levels and their balances are two factors affecting the growth of plant.  However, the responses of different crop cultivars to K/Mg ratios are less clear.  This study was aimed at assessing the different responses of tomato (Solanum Lycopersicum L.) cultivars to the different K/Mg supply ratios.  Three tomato cultivars (Zhongza 9 (ZZ), Gailiangmaofen (MF), and Jinpengchaoguan (JP)) were grown in pots with three different K+/Mg2+ ratios (4:0, 4:1 and 8:1, represented by K/Mg4:0, K/Mg4:1, and K/Mg8:1, respectively).  Compared with K/Mg4:1 treatment, the leaf chlorophyll content, net photosynthetic rate, and total biomass of tomato seedlings under K/Mg4:0 treatments were decreased by 69.7, 89.1, and 53.1%, respectively.  The Mg deficiency symptoms were observed when the Mg content in shoot became lower than 4 mg g–1 DW.  Compared with K/Mg4:1 treatment, total biomass of tomato seedlings of K/Mg8:1 treatment was decreased by 21.6%; the shoot and root Mg contents were decreased by 10.4 and 21.8%, respectively; and Mg uptake of tomato was reduced by 34.1%.  There were significant differences in biomass and Mg uptake for the three cultivars between the different K+/Mg2+ treatments.  The Mg uptake of the three different cultivars ranked as ZZ>JP>MF under Mg deficiency and high K condition.  In conclusion, the growth and Mg uptake and allocation of tomato were influenced significantly by imbalance K and Mg supply. JP and ZZ were the cultivars with the highest efficiency in Mg uptake. 
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4. Delivery of CatSper2 siRNA into Rat Sperms by Electroporation Repressed Ca2+ Influx During Sperm Hyperactivation
ZHANG Zhen, ZHOU Xuan, LI Hui-xia, CUI Qun-wei, YU Jing , WANG Gen-lin
Journal of Integrative Agriculture    2011, 10 (12): 1958-1967.   DOI: 10.1016/S1671-2927(11)60197-1
摘要1529)      PDF    收藏
CatSper is a unique Ca2+ channel-like protein family exclusively expressed in the testis and sperm, and plays important roles in sperm motility, capacitation, acrosome reaction and sperm-egg interactions. Here we studied the mechanism of regulation of CatSper2-dependent Ca2+ influx, extracellular and intracellular Ca2+ on sperm hyperactivated motility. The siRNA duplexes were transfected into the sperm cells by electroporation (EP) to silence the expression of CatSper2. The results for targeted disruption of CatSper2 showed the highest silence efficiency 77.7% (P<0.05), the hyperactivated sperm rate calculated by computer-assisted semen analysis (CASA) analysis of interferenced sperm was significantly lower 11.1% than the control 99.2%. Flow cytometry (FCM) detection of the intracellular Ca2+ concentration of interferenced sperm was 50 nmol L-1 higher than the normal. It was remarkably lower than hyperactivated sperm with 200-1 000 nmol L-1 (P<0.05). It was not sufficient to evoke hyperactivation. To trigger hyperactivation, the sperm were incubated in 50 μmol L-1 thimerosal or 5 mmol L-1 procaine, it sharply increased the intracellular Ca2+ via two different channels. CASA and FCM detection indicated that intracellular Ca2+ is required for initiating hyperactivation while extracellular Ca2+ is essential to maintain the process. We concluded that to mediate sperm hyperactivation, it is essential to inhibit Ca2+ peak present with targeted disruption of CatSper2 expression. This provides important prospective for further exploration of signal channel of sperm hyperactivated motility, potential factors for male infertility and provide further reference to the exploration of male contraceptive drug targets of male reproduction.
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