Diversity of nitrogen-fixing cyanobacteria under various ecosystems of Thailand: I. Morphology,physiology and genetic diversity

The diversity among 853 isolates of nitrogen-fixing cyanobacteria obtained from soil samples collected from different ecosystems including mountainous, forest and cultivated areas in the central, northern and northeastern regions of Thailand was examined. Most isolates showed slow growth rate and had filamentous, heterocystous cells. The percentage of heterocysts in the filaments of different isolates varied from 8.3 to 9.6. Only a few strains showed high nitrogen-fixing potential, while most of the strains exhibited low capacity for nitrogen fixation. Anabaena and Nostoc were the dominant genera among these isolates. One hundred and two isolates were randomly selected from this diverse collection to determine the extent of genetic diversity on the basis of DNA fingerprinting using the PCR method. Based on the PCR products obtained by using a combination of three primers, all strains could be distinguished from one another. When a subset of 45 isolates of Nostoc and a subset of 44 isolates of Anabaena were further analysed by PCR, a wide range of diversity was observed within each of these genera.     

Polyclonal antisera production by immunization with mixed cell antigens of different rhizobial species

Somatic antigens of Bradyrhizobium japonicum, Rhizobium sp. (Cicer arietinum) and Rhizobium sp. (Leucaena leucocephala) were prepared as standard, single-species type from cultured cells. Equal numbers of the cells of these rhizobia were then combined to obtain a mixed-rhizobial-species antigen preparation. Rabbits were immunized either with the standard, single-species type or with the mixed-rhizobial-species antigen preparations. The antisera developed from the mixed antigen immunization contained antibodies for all three rhizobial species, detectable at agglutination titres of over 800. The mixed-rhizobial-species antisera were made species specific by cross-absorption. The cross-absorbed and the mixed-rhizobial-species antisera were generally similar in quality for strain identification by agglutination, fluorescent antibodies, immunoblot and ELISA. A 66% reduction in cost was estimated for the production of antisera by immunization with mixed-rhizobial-species antigen.H.J. Hoben and P. Somasegaran are with the NifTAL Center and MIRCEN, University of Hawaii, 1000 Holomua Road, Paia, Maui, HI 96779-9744, USA: N. Boonkerd is with the School of Agricultural Technology, Suranaree University of Technology, University Avenue, Nakorn Racharsima, Thailand. Y.D. Gaur is with the Division of Microbiology, Indian Agricultural Research Institute, New Delhi-110012, India.     

Natural 15N abundance in shrub and tree legumes,Casuarina, and non N2 fixing plants in Thailand

The leaves and nodules from the shrub and tree legumes, particularly, Aeschynomene spp., Sesbania spp., Mimosa spp. and Leucaena spp., and Casuarina spp. and the leaves from neighbouring non-fixing plants were analyzed for their natural abundances of ¹⁵N ( ¹⁵N).The ¹⁵N in the leaves of non-fixing plants was +5.9% on average, whereas those from shrub legumes and Casuarina spp. were lower and close to the values of atmospheric N₂, suggesting the large contribution of N₂ fixation as the N source in these plants. The ¹⁵N values of the leaves from tree legumes except for Leucaena spp. were between the shrub legumes and non-fixing plants, which suggests that the fractional contribution of fixed N₂ in tree legumes may be smaller than that in the shrub legumes. Casuarina spp. was highly dependent on N₂ fixation. The ¹⁵N values of the nodules from most of the shrub legumes investigated were higher than those of the leaves.     

Salicylic acid-induced accumulation of biochemical components associated with resistance against Xanthomonas oryzae pv. oryzae in rice

Seed treatment and foliar sprays of salicylic acid (SA) provided protection in rice against bacterial leaf blight (BLB) caused by bacterial Xanthomonas oryzae pv. oryzae (Xoo). Treatment of rice with exogenous SA reduced disease severity by more than 38%. Superoxide anion production and hypersensitive response increased approximately 28% and 110% at 6 and 48?h after Xoo inoculation, respectively, for plants treated with SA. Moreover, the Xoo in treated rice plants grew more slowly, resulting in a population that was half of that observed in the control. Fourier transform infrared spectroscopy analysis revealed that the higher ratios of 1233/1517, 1467/1517, and 1735/1517?cm^?1 observed in treated rice suggested alteration of monomer composition of lignin and pectin in the rice cell wall. Exogenous SA-treated rice had more amide I β-sheet structure and lipids as shown by the peaks at 1629, 2851, and 1735?cm^?1. These biochemical changes of rice treated with SA and inoculated with Xoo were related to primed resistance of the rice plants to BLB disease.     

Cyanobacterial akinete induction and its application as biofertilizer for rice cultivation

Nostoc sp. VICCR1-1 was induced in order to form akinetes on the basis of nutrient modification. Phosphorus and iron were found to be the critical for akinete differentiation, especially when both elements were omitted. The number of akinete cells increased up to 20% when compared with culturing in BG11₀ medium (without N source). In addition, CaCl₂ played a role in heterocyst differentiation, and was able to induce heterocyst ranging between 30% and 46%. In order to prepare akinetes as inoculum, the dried form of akinetes was prepared by mixing it with montmorillonite clay. The inoculum with the amount of 2.8 × 10⁶ cells m⁻² was applied to rice (Oryza sativa) fields. After harvesting, the grain yields from chemical N fertilizer, vegetative cells, and akinete inoculum treatments were not significantly different. To monitor the persistence of Nostoc sp. VICCR1-1 after harvesting, the most probable number-denaturing gradient gel electrophoresis technique using 16S rRNA gene was employed. The results indicated that the remaining population is at 2.5 × 10⁵ and 1.62 × 10⁶ cells m⁻² in treatments supplied with vegetative cells and akinete inocula, respectively. Akinete induction might be one of the appropriate approaches for producing cyanobacterial inoculum.     

Mnesithea thailandica a new species of Poaceae from Thailand

Mnesithea thailandica, a new species of Poaceae subtribe Rottboelliinae from Thailand, is described and illustrated.     

Effects of N-serve on soybeans and soil nitrogen transformations

Summary Plant dry weight, total N, and total Ca was increased at 0.1 and 1 ppm N-serve. At greater 10 ppm the plants showed visual symptoms of a stunted growth, stem elongation, flowers, and pods failed to form or were aborted, young leaves were curled, and roots were club shaped with many branches. These symptoms were increasingly evident with increasing N-serve application rates. The reason was attributed to an auxin effect. Dry wt and total N in the plant was less than the control at the higher N-serve applications. There was little effect on nitrogenase activity at less than 10 ppm N-serve. Nodulation tended to increase at 0.1 and 1 ppm N-serve.Nitrification was inhibited up to 104 days at 20 ppm N-serve. The soil pH of the high N-serve rates was decreased at 104 days probably due to nitrification. Generally there were little detectable differences among treatments in soil organic N. The average soil organic N from 0 to 104 days decreased by 0.01%. Average increase in total N within each pot at harvest was equivalent to about 138 kg N/ha.