Incompatible Nodulation of <i>Bradyrhizobium elkanii</i>Strains BLY3-8 and BLY6-1 with <i>Rj</i>3 Gene-Harboring Soybean Cultivars

Bradyrhizobia are known symbiotic partners of soybean. However, some soybean cultivars restrict nodulation by some Bradyrhizobium bacterial strains. These restrictions are related to compatibility between the Rj genes of soybean cultivars and nodulation types of inoculated bacteria. The objective of this study was to determine nodulation incompatibility of Type B strains with Rj3 soybean cultivars. Newly isolated B. elkanii strains BLY3-8 and BLY6-1 from Myanmar and specific strain Bradyrhizobium elkanii USDA33, which are incompatible with Rj3 soybean cultivars, and B. japonicum USDA110 were used as inoculants to check compatibility or incompatibility with Rj3 soybean cultivars. Nitrogen fixation activity was measured by the acetylene reduction method. Ethylene concentration (reduction of acetylene) was determined by flame ionization gas chromatography. According to the inoculation test results, USDA110 was compatible with all soybean cultivars because it formed effective nodules (Figure S1 in Appendix) and possessed nitrogenase activity. Similarly, B. elkanii strains BLY3-8, BLY6-1, and USDA33 were highly compatible with non-Rj and Rj4-gene harboring soybean cultivars because they had the ability to form functional nodules and possessed nitrogenase activity. Inversely, BLY3-8, BLY6-1, and USDA33 were incompatible with Rj3 soybean cultivars because they produced ineffective nodules. Consequently, the ratio of ineffective nodule number to total nodule number was >0.5. Therefore, nodule formation by the newly isolated B. elkanii strains BLY3-8 and BLY6-1 was restricted by the Rj3 soybean cultivars potentially making them useful as specific strains to detect the Rj3 gene in soybean cultivars.

大豆埃氏慢生根瘤菌Y63-1菌株培养基的筛选与优化

为了促进广谱高效大豆埃氏慢生根瘤菌Y63-1菌株的高效生产与推广应用,本研究对其培养基进行了筛选与优化。首先比较了Y63-1在YMA、TY、SM、PA和BSE 5种根瘤菌基本培养基中的生长速度,结果表明Y63-1在TY基本培养基中生长最快。以TY为基本培养基进行单因素碳源及无机盐利用试验,结果表明葡萄糖是Y63-1生长的最佳碳源,CaCl_(2)为必要的培养基成分,Rh微量元素对菌株的生长有很大的促进作用。进一步对蛋白胨、葡萄糖、酵母粉及Rh微量元素4种组分进行正交优化,获得了适宜Y63-1生长的最佳培养基,配方(1 L)为:8 g蛋白胨、10 g葡萄糖、3 g酵母粉、0.1 g CaCl_(2)·6H_(2)O、3 mL Rh微量元素,pH7.0。此培养基也能显著提升USDA110的生长速率,可以广泛应用于慢生根瘤菌菌剂的大规模生产。

根瘤菌与烯效唑互作对大豆生长及固氮的影响

以根韦氏中华根瘤菌(Sinorhizobium fredii)和埃尔坎慢生根瘤菌(Bradyrhizobium elkanii)为研究材料,以0.001~100 mg·L^(-1)浓度的烯效唑培养两种根瘤菌,探究烯效唑对根瘤菌生长的影响。结果表明:供试烯效唑浓度范围内大于10 mg·L^(-1)时抑制根瘤菌数量,烯效唑浓度为1 mg·L^(-1)时对根瘤菌数量具有促进作用。大豆以1 mg·L^(-1)浓度烯效唑浸种,每2.5 kg土壤拌有30 m L对数生长期的根瘤菌菌液,在花荚期测定大豆叶片与根系的SOD、POD、CAT酶活力、丙二醛含量(MDA)及大豆根系酰脲含量,确定根瘤菌与烯效唑互作对大豆生长及共生固氮的影响。结果显示:与根瘤菌处理相比,根瘤菌与烯效唑互作可降低大豆植株中MDA含量达40%~60%,提高大豆根系酰脲含量达5%~8.4%,但对测定的3种酶活性影响不显著。试验结果证实了根瘤菌与烯效唑互作可提高大豆固氮能力,降低MDA含量。