枯、黄萎病水培接菌方法室内抗病鉴定方法

1、棉花枯、黄萎病室内抗病鉴定方法棉种催芽后种在纯蛭石中，三天左右待子叶平展后拔出，此时未长出侧根， 只有主根，易于拔出且不易造成对根的损伤。用打孔器在黑色泡沫板上打取直径 约2cm的孔洞，将打好洞的泡沫板放入装有hoagland营养液的塑料盘中，使泡 沫板刚好卡在塑料盘口，然后用海绵条轻轻缠绕发芽棉种23圈，然后将其塞入 泡沫板的孔洞中，根芽向下浸入水中。放置25。（2的培养室，光照16h,黑暗8h 培养。W-80保存的菌株经PDA平板活化3-4天，然后用查比克培养基振荡培养 3-4天，待有两片真叶时接菌，接菌浓度1义IO,。将棉苗根浸泡在菌液中24h,然后再换成营养液，两周后开始发病。下图是水

2、培棉苗。依V) n下面是郭慧珊老师那边水培接种的方法Cotton plants (cv. Xinluzao NO. 16) were used in infection assays to evaluate the effect of V.dahliae isolate V592 mutation on virulence. Mutants generated from V592 were evaluated for their virulence on cotton (cv. Xinluzao NO. 16), using our laboratory unimpaired root dip

3、-inoculation method”. To prepare inocula, fungal cultures grown for 5 days in the liquid Czapek-Dox medium were passed through several layers of cheesecloth (to remove mycelia), and the conidial concentration was adjusted to approximately IxlO7 conidia per ml. 12 Seedlings per pot were planted in MS

4、 liquid medium (Murashige and Skoog medium) in an environmentally controlled growth room at 261, 60-70% relative humidity, on a 16-h light/8-h dark cycle, for about two weeks. At the third true leaf stage, the seedlings were then inoculated by immersing their roots into the conidial suspension for 5

5、0 min. The seedlings were then put back to the 1/10 MS liquid medium. Disease progress was recorded over time for two months of experiment period. Disease severity was counted by the percent of leaves that showed wilting symptom at each time point. Infection assay for each mutant colony was repeated

6、 at least three times.H同7番低m (UWrlUWOI 吠c.4i47-20“WIO 合格 OOOr% DCOS%卬.wn10010618丙三醇（甘油）Glycerol分析纯ARC凡o,J 澄清.粘璜液体、无, : 92.09 91-5 J20160309.ItttSOGOO，： 2I队内凡3WM砂 、1V1/ L /(g)KNOj0.3838190MgSO40.033.8/7.4(MgSO-i 7112O)19/37KH2PCh0.0253.417CaCh0.066.6/8.8(CaCh 2 I hO)33/44单独溶解，最。微量元素母液配制药品质量浓度(g/L)备注5倍母液IHsBOs3.1加热助溶MnSO4 4H2O (或MnSO4 H2O)11.15 (或 8.45)ZnSO4 7HO4.320倍母液IIC0CI2 6H2O0.05CuSO4 5H2O0.05单独溶解，最后加入KI1.66NaMO4 2H2O0.5取上述母液1 200ml,母液II50mL加蒸储水定容至IL。铁盐母液配制