First report of little leaf disease associated with phytoplasma on sugar beet (Betavulgaris L. subsp. vulgaris var. altissima Doll) in India

Sugar beet (Beta vulgaris L. subsp. vulgaris var. altissima) is a biennial, sugar-producing tuber crop grown in different parts of the world. Diseased sugar beets with symptoms of little leaf were observed during 2008 in Tamil Nadu, India. Phytoplasmas were detected in symptomatic leaves of three separate plants using PCR with primer pair fU5/rU3, which amplify an 880-bp fragment of the 16S rRNA region of phytoplasmas. The nucleotide sequence analysis of a 766-bp fragment had 100% identity among the sequence from the three plants (GQ184437) and 96% nucleotide sequence identity with 16S ribosomal RNA gene sequences of eggplant phyllody phytoplasma.     

Cucumber performance is improved by inoculation with plant growth-promoting microorganisms

We investigated the effects of inoculation of Rhodobacter sphaeroides, Lactobacillus plantarum, and Saccharomyces cerevisiae on cucumber plant growth promotion and on the contents of plant hormones, amino acids, and mineral nutrients. We showed that treatment with all three bio-inoculants significantly increased the shoot length, root length, shoot fresh weight, shoot dry weight, and chlorophyll content, via secretion of indole acetic acid and/or organic acids. Inoculation with R. sphaeroides had more favorable effect on plant growth than did inoculation with L. plantarum or S. cerevisiae, by significantly enhancing the gibberellin and reducing the abscisic acid contents. The results of amino acid analysis revealed that inoculation with R. sphaeroides, L. plantarum, and S. cerevisiae generally increased the contents of 17 amino acids, namely, aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine, and proline. With the exception of cysteine, all these amino acids were present in higher concentrations in plants inoculated with R. sphaeroides than in control plants or in plants inoculated with L. plantarum and S. cerevisiae. Furthermore, inoculation with R. sphaeroides significantly increased the calcium, potassium, magnesium, and phosphate contents. Our results suggest that the use of R. sphaeroides, L. plantarum, and S. cerevisiae in agricultural fields can improve plant growth. Moreover, inoculation of cucumber plants with R. sphaeroides regulates plant functional metabolites, thereby promoting plant growth.     

A significant proportion of indigenous rhizobia from India associated with soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.) distinctly belong to <Emphasis Type="Italic">Bradyrhizobium</Emphasis> and <Emphasis Type="Italic">Ensifer</Emphasis> genera

The diversity among 269 rhizobia isolated from naturally occurring root nodules of soybean collected from two different agro-ecological regions of India, based on RFLP and sequences of the intergenic spacer (IGS) between the 16S and 23S rRNA genes, growth rate, and indole acetic acid production, revealed their significant, site-dependent genomic diversity. Among these bacteria, nine IGS genotypes were identified with two endonucleases. They were distributed into five divergent lineages by sequence analysis of each IGS representative strain, i.e., (1) comprising IGS genotypes I, II, III, and reference Bradyrhizobium yuanmingense; (2) with genotype IV and strains of unclassified bradyrhizobia genomic species; (3) including genotypes V, VI, and Bradyrhizobium liaoningense; (4) with IGS genotype VII and Bradyrhizobium elkanii strains; and (5) comprising IGS genotypes VIII, IX, and different Ensifer genus bacteria. Host-specificity test revealed that all rhizobia-nodulated soybean and cowpea and only part of them formed nodules on Arachis hypogeae and Cajanus cajan. The great diversity of soybean nodulators observed in this study emphasises that Indian soil is an important reservoir of nitrogen-fixing rhizobia.     

Isozyme pattern of lactate and malate dehydrogenases of Gastrothylax crumenifer (Trematoda: Amphistomatidae) from different hosts

Isozyme pattern of lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) of Gastrothylax crumenifer from sheep, goat and buffalo was studied using polyacrylamide gel electrophoresis. LDH of G. crumenifer from buffalo, goat and sheep consists of four fractions, three fractions and two fractions, respectively. The parasite from buffalo shows two fractions of MDH, whereas those from goats or sheep show only a single fraction. The significance of these results is discussed.     

Tissue culture and <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated genetic transformation studies in four commercially important indica rice cultivars

This research was undertaken to find an efficient tissue culture system and Agrobacterium-mediated genetic transformation method for recalcitrant indica rice cultivars. For this, mature seeds of commercially important indica rice varieties, ASD16, ADT43, IR 64, and Pusa Basmati were cultured on MS and N6 medium supplemented with 2 mg l^-1 2, 4-D + 30 g l^-1 sucrose. The calli grown in N6 medium showed better friability and embryogenic response. Out of the four varieties tested, ASD16 and IR64 showed better callusing and embryogenic capacity as compared to ADT43 and Pusa Basmati. For genetic transformation studies, embryogenic calli of all the cultivars were co-cultivated with the Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pCambia 1305.1 with GUS gene. GUS assay was performed for the putative transformed calli and its activity was found to be qualitatively higher in ASD16 and IR64 than the other two varieties. The best responsive ASD16 transformed calli was regenerated and the putative transgenic lines were regenerated. ASD16 transformed calli were confirmed by GUS assay. PCR analysis confirmed the presence of both GUS and HPT genes in ASD16 transgenic lines.     

Characterization of esterase isozymes of Raillietina tetragona (Molin, 1858) (Cestoda)

Soluble esterase isozymes from Raillietina tetragona were separated in polyacrylamide gel electrophoresis, which revealed the presence of at least four fractions, possibly representing different esterases. The differences in the inhibitory effect of various chemicals on these fractions reveal them to be four types of esterases, namely: arylesterase, carboxylesterase, acetylesterase and cholinesterase.     

Mutant prevention concentration,pharmacokinetic–pharmacodynamic integration,and modeling of enrofloxacin data established in diseased buffalo calves

The pharmacokinetic–pharmacodynamic (PK/PD) modeling of enrofloxacin data using mutant prevention concentration (MPC) of enrofloxacin was conducted in febrile buffalo calves to optimize dosage regimen and to prevent the emergence of antimicrobial resistance. The serum peak concentration (C_max), terminal half‐life (t_1/2K₁₀₎, apparent volume of distribution (Vd_(area)/F), and mean residence time (MRT) of enrofloxacin were 1.40 ± 0.27 μg/mL, 7.96 ± 0.86 h, 7.74 ± 1.26 L/kg, and 11.57 ± 1.01 h, respectively, following drug administration at dosage 12 mg/kg by intramuscular route. The minimum inhibitory concentration (MIC), minimum bactericidal concentration, and MPC of enrofloxacin against Pasteurella multocida were 0.055, 0.060, and 1.45 μg/mL, respectively. Modeling of ex vivo growth inhibition data to the sigmoid E_max equation provided AUC_24 h/MIC values to produce effects of bacteriostatic (33 h), bactericidal (39 h), and bacterial eradication (41 h). The estimated daily dosage of enrofloxacin in febrile buffalo calves was 3.5 and 8.4 mg/kg against P. multocida/pathogens having MIC₉₀ ≤0.125 and 0.30 μg/mL, respectively, based on the determined AUC_24 h / MIC values by modeling PK/PD data. The lipopolysaccharide‐induced fever had no direct effect on the antibacterial activity of the enrofloxacin and alterations in PK of the drug, and its metabolite will be beneficial for its use to treat infectious diseases caused by sensitive pathogens in buffalo species. In addition, in vitro MPC data in conjunction with in vivo PK data indicated that clinically it would be easier to eradicate less susceptible strains of P. multocida in diseased calves.     

Association between root growth angle and root length density of a near-isogenic line of IR64 rice with DEEPER ROOTING 1 under different levels of soil compaction

DEEPER ROOTING 1 (DRO1) of rice controls the gravitropic response of root growth angle. In order to clarify the effects of DRO1 on root growth angle and root length density under different soil resistance to penetration, and to quantify the relationship between root growth angle and root length density, we assessed the root growth of Dro1-NIL (a near-isogenic line homozygous for the Kinandang Patong allele of DRO1 in the IR64 background) under upland Andosol field conditions in Japan in 2013 and 2014. The trial included three levels of soil compaction (none, moderate, and hard). Root length density at a depth of 30 to 60 cm was largest in Kinandang Patong, followed by Dro1-NIL, and was least in IR64 in both years and in all compaction treatments. Root length density at this depth decreased with hard compaction (to 70% of control) and increased with moderate compaction (to 135%). The number of roots with a deep angle (i.e. 45° to 90° from the horizontal) measured by the basket method was similar at maximum tillering and maturity stages, and its value as a proportion of the total number of roots was strongly correlated with the root length density at 30 to 60 cm in both years, which demonstrates the importance of a deep root angle for the development of deep roots. Dro1-NIL had a higher proportion of deep roots than IR64, but the difference was small under hard compaction, with a significant genotype × compaction interaction.     

STS and microsatellite marker-assisted selection for bacterial blight resistance and waxy genes in rice, Oryza sativa L.

DNA marker-assisted selection was employed to select Xa-21 bacterial blight resistance and waxy (Wx) genes. Genotypes with both genes were selected from four F₂ populations involving indica × indica, indica × intermediate and japonica × indica crosses. With the assistance of PCR marker, 13 true breeding lines carrying Xa-21 were identified from F₂ generation of IRBB 21 × G 11353 cross. Similarly ten, eleven and fifty two plants having Xa-21 gene were isolated from G 3005-4-1 × IRBB 21, IRBB 21 × HJX 74 and IRBB 21 × SY 2crosses respectively. The lines with Wx gene in both homozygous and heterozygous state were also scored from the above crosses. Twenty plants having both Xa-21 and Wx loci in homozygous state were identified. DNA-based progeny testing was conducted to ensure the selection of homozygous lines for Xa-21 and Wx genes. Finally,twenty true breeding lines with high amylose content and Xa-21 gene were isolated from four crosses. These homozygous lines are phenotypically superior and resistant to Chinese race 5 of the bacterial blight pathogen. Fifty-six germplasm sources were surveyed for PCR polymorphism in order to facilitate future PCR-based marker assisted transfer of bacterial blight resistance genes xa-5, xa-13 and Xa-21 to any desired varietal background which will be useful for selection of parents in breeding programmes. This revised version was published online in August 2006 with corrections to the Cover Date.     

The host background of rice influences the resistance expression of a three genes pyramid (xa5 + xa13 + Xa21) to bacterial blight (Xanthomonas oryzae pv. oryzae) pathotypes of Indian mainland and Bay islands

Bacterial blight (BB) is a major disease of rice for which host resistance is the only effective solution. The three genes pyramid xa5 + xa13 + Xa21 is recently the most utilized combination for developing resistant varieties through marker‐assisted breeding. Our study was carried out to elicit the detailed response of twenty lines possessing these three genes in five genetic backgrounds to twelve diverse BB pathotypes in India. The lines developed from ADT 47 variety showed incomplete resistance to most of the pathotypes, whereas susceptibility varied from 8.3% to 16.6% in ADT 43 and IR24, respectively. However, in IMP ASD16/60 and Improved Samba Mahsuri, complete resistance against all pathotypes was observed. The overall results confirmed that genetic background plays crucial role for the effective expression of xa5 + xa13 + Xa21 combination. Molecular studies did not reveal correlation between origin of pathotypes and their virulence potential. It is suggested to deploy Improved Samba Mahsuri, IMP ASD 16/60 and AD1306 varieties in the bacterial blight prone areas or use them as donors for realizing wider and durable resistance.