Low-Density Co-Inoculation with Bradyrhizobium japonicum SAY3-7 and Streptomyces griseoflavus P4 Promotes Plant Growth and Nitrogen Fixation in Soybean Cultivars

Inoculation density has a marked effect on nodulation and N fixation in soybean (Glycine max L.). Therefore, we conducted this study to determine the optimal inoculation density of Bradyhizobium japonicum SAY3-7 (SAY3-7) and Streptomyces griseoflavus P4 (P4) for plant growth, nodulation, and N fixation, and to investigate the effect of co-inoculation on selected soybean cultivars, using the optimal inoculation density. Nitrogen fixation, in terms of an acetylene reduction activity value, was measured using a flame ionization gas chromatograph. In this study, low-density single inoculation with P4 (105 or 106 cells mL-1) was associated with the highest plant biomass, compared with normal- and high-density single inoculation with P4 (107 or 108 cells mL-1). Moreover, low-density single or co-inoculations with SAY3-7 and/or P4 produced the highest nodule biomass and highest nitrogenase activity, compared with single or dual inoculation at other inoculation densities. Therefore, we evaluated low-density co-inoculation with P4 and SAY3-7, at the rate of 105 cells mL-1, on selected soybean cultivars. Low-density co-inoculation increased the plant biomass, compared with un-inoculated plants. The effects of single and co-inoculation on nodulation did not differ significantly for any of the cultivars, except “Yezin-9” in the first experiment and “Shan Seine” in the second experiment. Low-density inoculation with both bacteria increased N fixation by 15% - 75% for seven of the cultivars in the first experiment and by 15% - 39% for three of the cultivars in the second experiment, compared with single inoculation with SAY3-7. Based on the overall results, we concluded that low-density co-inoculation with P4 and SAY3-7 gave improved plant growth and N fixation.

Effects of Integrated Organic and Inorganic Fertilizers on Yield and Growth Parameters of Rice Varieties

We investigated the effects of integrated organic and inorganic fertilizers on the growth and yield of indica rice variety Manawthukha and japonica rice variety Genkitsukushi.In a split-plot design,the two rice varieties were assigned as main plot factors,and the integrated treatments were the subplot factors,including no-N fertilizer(N0),50%chemical fertilizer(CF)(CF50),100%CF(CF100),50%CF+50%poultry manure(PM)(CF50PM50),50%CF+50%cow manure(CM)(CF50CM50),and 50%CF+50%compost(CP)(CF50CP50).CF100 was equivalent to N at 85 kg/hm2.Manure was applied based on the estimated mineralizable nitrogen(EMN)level,which is dependent on total N(%)of each manure type.Manawthukha rice plants were taller with higher tiller number and dry matter content.However,higher soil-plant analysis development(SPAD)values were measured in Genkitsukushi throughout the crop growth period,resulting in higher seed-setting rate(%)and greater yield.At the same N level,CF50PM50 application in both rice varieties resulted in higher SPAD values,plant height and tiller number than CF100.CF50PM50 containing total N more than 4%supplied synchronized N for the demands of the rice plants,resulting in maximum dry matter,yield and yield components.CF50CM50 and CF50CP50 treatments containing total N less than 4%resulted in lower yields which were similar to CF100.These results indicated that integrating organic and inorganic fertilizers enhanced growth parameters and yields of Manawthukha and Genkitsukushi,while reducing the dose of chemical fertilizer.

Effects of Fermented Nori (<i>Pyropia yezoensis</i>) Liquid Fertilizer on Plant Growth Characteristics and Nutrient Content of Komatsuna (<i>Brassica rapa</i>L. var. Wakana Komatsuna) Cultivated in Vermiculite

We conducted plant growth experiments in microbe-free vermiculite to study the effects of four types of fermented seaweed liquid fertilizer (SLF) made from nori (Pyropia yezoensis) seaweed on the germination, plant growth characteristics, SPAD value, and nutrient content and uptake of komatsuna (Brassica rapa L. var. wakana komatsuna). The four types of fermented nori SLF used in this study were prepared by anaerobic fermentation of unwashed nori (SLF1), aerobic fermentation of unwashed nori (SLF2), anaerobic fermentation of washed nori (SLF3), and aerobic fermentation of washed nori (SLF4). Komatsuna seeds treated with 200-, 300-, and 400-fold dilutions of SLFs exhibited improved relative germination ratios (RGRs) at 3 and 4 days after sowing (DAS). At 4 DAS, the RGRs of seeds treated with 10-, 100-, 200-, 300-, and 400-fold SLF dilutions showed no differential effect. Seeds treated with undiluted SLFs did not germinate by 4 DAS. SLF1 may promote komatsuna seed germination. The nitrogen (N), calcium, magnesium, sodium (Na), and iodine (I) contents of plants treated with SLF1 were significantly increased relative to plants treated with the other SLFs. Moreover, the I and Na contents of plants were significantly increased by foliar spray application of different dilutions of SLF1. However, SLF treatment markedly reduced the shoot dry weight compared with 1/2-strength modified Hoagland nutrient (MHN) solution, although the same amounts of N and K were applied. SPAD values of the plants treated with SLFs were significantly higher than those of plants treated with MHN. Foliar treatment with SLFs had no significant effect on plant growth, SPAD value, or uptake of nutrients (except Na) relative to the control, but the I content was increased. Plants treated with SLF1 and SLF2 exhibited the highest Na uptake. Foliar spray treatments with SLF1 resulted in the highest I contents in plants. Based on our results, SLF1 is suitable for use as a liquid fertilizer to promote germination and increase nutrient content in komatsuna. These results need to be followed up in soil experiments in the presence of microbes in the rhizosphere.

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.

Determining Nodulation Regulatory (Rj) Genes of Myanmar Soybean Cultivars and Their Symbiotic Effectiveness with <i>Bradyrhizobium japonicum</i>USDA110

Soybean (Glycine max L.) plays an essential role in human nutrition as a protein source, and in plant nutrition as a N source. The rate of N fixation varies depending on the cultivars and compatibility between the inoculated Rhizobium strain and the host cultivar. Characterizing the nodulation regulatory (Rj) genes is necessary to determine the compatibility of cultivars and Rhizobium strains. Rj genes were previously identified based on inoculation tests and PCR analyses. The six cultivars Yezin-3, Yezin-7, Yezin-11, Shan Seine (Local), Madaya (Local), and Hinthada (Local) were identified as harboring the Rj4 gene. Two cultivars, Yezin-6 and Yezin-8, were classified as non-Rj-gene harboring. Two other cultivars, Yezin-9 and Yezin-10, were identified as Rj3- and Rj2Rj3-gene harboring, respectively. Ours is the first report on Rj3- and Rj2Rj3-gene harboring cultivars in Myanmar. We evaluated Myanmar soybean cultivars for symbiotic effectiveness, relying on the standard strain Bradyrhizobium japonicum USDA110. In our first experiment, the soybean cultivar Yezin-11 (Rj4) showed the highest N fixing potential. Based on their potential for fixing N and nodulation, the top six soybean cultivars were Yezin-11 (Rj4), Yezin-9 (Rj3), Yezin-6 (non-Rj), Yezin-8 (non-Rj), Yezin-3 (Rj4) and Yezin-10 (Rj2Rj3). These cultivars were selected for a second experiment, which revealed that the N fixation, nodulation, and plant growth of Yezin-11 (Rj4) *Corresponding author. A. Z. Htwe et al. 2800 were superior to the other cultivars. We conclude that Yezin-11 (Rj4) is the most efficient cultivar for nodulation and N fixation when inoculated with B. japonicum USDA110.

Effects of Fermented Nori (<i>Pyropia yezoensis</i>) Seaweed Liquid Fertilizers on Growth Characteristics, Nutrient Uptake, and Iodine Content of Komatsuna (<i>Brassica rapa</i>L.) Cultivated in Soil

We investigated the effects of fermented nori (Pyropia yezoensis) liquid fertilizers on plant growth, soil plant analysis development (SPAD) values, and nutrient uptake of komatsuna (Brassica rapa L. var. wakana komatsuna) plants. The four types of fermented nori seaweed liquid fertilizers (SLFs) evaluated in this study were prepared by anaerobic fermentation of unwashed nori (SLF1), aerobic fermentation of unwashed nori (SLF2), anaerobic fermentation of washed nori (SLF3), and aerobic fermentation of washed nori (SLF4). In Experiment 1, the highest plant growth, SPAD, and nutrient uptake values were obtained from treatment with SLF2 and SLF4. There were no significant differences between the effects of basal and foliar application of SLFs, except for iodine (I) content;plants treated with SLF1 had the highest I content. In Experiment 2, plant growth and nutrient uptake decreased with higher concentrations of SLFs. Plants treated with 25% SLF2 + 75% chemical fertilizer (CF) or 25% SLF4 + 75% CF exhibited significantly higher plant growth and nutrient uptake. The highest I content resulted from treatment with 75% SLF1 + 25% CF or 100% SLF1. Taken together, our results showed that 25% SLF + 75% CF produced the best plant growth characteristics, nutrient uptake, and I content relative to the controls. Therefore, basal application of these liquid organic fertilizers can be used to increase productivity, nutrient uptake, and I content and to reduce nitrate-nitrogen content in komatsuna production.

Phylogenetic Diversity and Evaluation the Effectiveness of Indigenous <i>Bradyrhizobium</i>Strains for Myanmar Black Gram (<i>Vigna mungo</i>L. Hepper) Cultivars

Black gram (Vigna mungo L. Hepper) is one of the main leguminous crops that provide chief source of food. Several Bradyrhizobium species are able to induce effective nodules in black gram cultivars. In the present study, we characterized forty isolates of indigenous black gram bradyrhizobia from Myanmar based on the sequence analysis of the bacterial 16S rRNA gene. The sequence analysis confirmed that all isolates were categorized and identified as the genus Bradyrhizobium and they were conspecific with B. elkanii, B. sp., B. liaoningense, B. japonicum and B. yunamingense. Almost all the collected isolates from major black gram growing regions of Nyaunglebin Bago Regio, Chaungzon Mon State, Sittwe Rakhine State, Danubyu Ayeyarwady Region and Launglon Tanintharyi Region were identified as B. liaoningense. At Danubyu Ayeyarwady Region and Pyinmanar Nay Pyi Taw Region, most of the strains were identified as B. japonicum. On the other hand, more or less all the isolates from Launglon Tanintharyi Region and Hpa-an Kayin State were related to B. elkanii. However, all B. sp. strains were found in Salingyi Sagaing Region black gram growing region. This is the first report describing Bradyrhizobium strains that were isolated from soil samples of major black gram growing areas in Myanmar. Evaluation of the effectiveness of Myanmar Bradyrhizobim strains isolated from soil samples of major black gram growing areas of Myanmar for plant growth and nitrogen fixation was studied in pot experiments with completely randomized design and three replicates. The nodule dry weight, shoot dry weight and acetylene reduction activity of the plant inoculated with Bradyrhizobium elkanii LauBG38 were significantly higher in ARA per plant, nodule and shoot dry weights than the other tested isolates in both Yezin-4 and Yezin-7 black gram varieties. We expect that Myanmar Bradyrhizobium elkanii LauBG38 will be able to use as Biofertilizer for black gram cultivars.

Dry Weight Accumulation, Root Plasticity, and Stomatal Conductance in Rice (<i>Oryza sativa</i>L.) Varieties under Drought Stress and Re-Watering Conditions

Drought is one of the main factors limiting rice (Oryza sativa L.) productivity and has become an increasingly severe problem in many regions worldwide. Establishing breeding programs to develop new drought-tolerant varieties requires an understanding of the effect of drought on rice plants and the mechanisms of drought tolerance in rice. We conducted a pot experiment to explore growth characteristics, root plasticity, and stomatal conductance in six rice varieties (DA8, Malagkit Pirurutong, Thierno Bande, Pate Blanc MN1, Kinandang Patong, and Moroberekan) in response to different drought stress and re-watering conditions. Drought stress significantly depressed plant growth, root size, and stomatal conductance in all experimental varieties. These negative effects depended on both the variety and the severity of the drought stress treatment. Under moderate drought stress (10 days after drought treatment), growth was less influenced in roots than in shoots. In contrast, there was an opposite trend under severe drought stress (15 days after drought treatment), with growth being more severely affected in roots than in shoots. Rice plants recovered from drought stress in terms of dry matter accumulation, root size, and stomatal conductance after re-watering;however, the recovery pattern differed among varieties. DA8 exhibited the highest dry weight accumulation and root size (root length, root surface area, root volume, fine root length, and thick root length) under well-watered, drought stress, and re-watering conditions. Kinandang Patong showed the highest recovery ability in dry matter accumulation, root length, root surface area, and stomatal conductance after re-watering. Malagkit Pirurutong expressed the poorest recovery ability in dry matter accumulation after re-watering. These three varieties might be selected for further experiments focusing on the mechanisms of drought tolerance and recovery ability in rice.