Alleviation of drought stress in maize (Zea mays L.) by using endogenous endophyte Bacillus subtilis in North West Himalayas

ABSTRACT

In the present study, we reported the isolation and molecular characterisation of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing endophyte and its effect on alleviating drought stress in maize. Ten morphologically distinct indigenous plant growth-promoting rhizobacteria (PGPR) from maize roots and rhizosphere were screened for multiple plant growth-promoting (PGP) traits, and of ten, two strains (DHK and B1N1) possessing maximum PGP traits were evaluated at three water stress regimes in maize: (i) 80% field capacity (FC), (ii) 60% FC, and (iii) 40% FC for 45 days, starting 15 days after sowing to the maturity. Inoculation of DHK strain with maize at 80% FC induced a significant increase in plant biomass (root biomass, 86.67%; shoot biomass, 111.63%) together with decreased reactive oxygen species and increased activity of antioxidant enzymes (superoxide dismutase, 14.11?U/g fresh weight; peroxidase, 1.14?U/g fresh weight and catalase; 14.91?U/g fresh weight) compared to uninoculated control at same water regime (80% FC). The derivative strain also significantly accrued nutrients in maize and enhanced total chlorophyll and amino acid content in comparison with non-inoculated plants. 16S rDNA analysis of DHK strain revealed its lineage to Bacillus subtilis. Present investigations demonstrated the potential of the bacterial partner in alleviating drought stress in maize.     

Induction of somatic embryogenesis and analysis of genetic fidelity of in vitro<Emphasis Type="Italic">-</Emphasis>derived plantlets of <Emphasis Type="Italic">Bambusa nutans</Emphasis> Wall., using AFLP markers

Bambusa nutans Wall., is an evergreen, perennial, and multipurpose bamboo having strong culms, which are largely used for construction, scaffolding, craft purposes, pulp, and paper industry. Multiple shoots from nodal segments (3–4 cm) of young branches of mature culms were established in Murashige and Skoog (1962) (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BAP) (1.0–6.0 mg l⁻¹) or in combination with α-naphthaleneacetic acid (NAA) (0.5–1.0 mg l⁻¹) or kinetin (Kn) (1.0–2.0 mg l⁻¹). February–March and December were found to be the best seasons for culture establishments. Maximum shoots were achieved on MS medium fortified with BAP (2.0 mg l⁻¹). Embryogenic callus (slightly greenish compact, globular, and slow growing) was initiated from the base of severed sprouted buds in 2–3 subsequent subcultures on MS medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) (5.0 mg l⁻¹) under dark incubations. Maturation and germination of well-organized somatic embryos was achieved on MS medium containing BAP and 2,4-D (1.0 mg l⁻¹ each) with 20.0 mg l⁻¹ ascorbic acid. Full-strength MS medium supplemented with 2% glucose favored further development of proliferated somatic embryos into plantlets. Genetic variations of field-established B. nutans plants regenerated through tissue cultures were assessed by amplified fragment length polymorphism (AFLP) analysis using 6 primer combinations. Four hundred and seven scorable fragments were amplified, of which 402 (98.8%) have recorded conservation at various morphogenetic stages leading to plantlets regeneration, therefore, revealed a high level of genetic stability.     

In vitro Release of Ovarian Progesterone is Decreased During the Oestrous Cycle and Pregnancy of Swine with Obesity/Leptin Resistance

Previous studies indicate that reproductive prolificacy of obese swine breeds is markedly influenced by embryo losses in early pregnancy. In such period, adequate secretion of progesterone (P4) by the ovary is essential for pregnancy success. This study analyses the luteal functionality during the oestrous cycle and early pregnancy of Iberian sows and Large White x Landrace females, in terms of P4 secretion after in vitro culture of luteal tissue stimulated or not with luteinizing hormone (LH). The secretion of progesterone (expressed in ng/mg of luteal tissue or ng/mgLT) of the corpora lutea of obese Iberian swine was always hampered when compared to lean genotypes, either during early oestrous cycle (110.7 ± 37.8 vs 259.7 ± 10.2 ng/mgLT; p < 0.0001), late oestrous cycle (49.0 ± 3.5 vs 75.92 ± 7.14 ng/mgLT; p < 0.0001) or early pregnancy (38.4 ± 2.1 vs 70.7 ± 5.3 ng/mgLT; p < 0.0001). The differences in basal P4 secretion remained after stimulation with LH. Finally, P4 secretion during early pregnancy of Iberian sows decreased with age and, hence, with obesity features (46.6 ± 4.2 vs 65.5 ± 4.8 ng/mgLT; p < 0.001). In conclusion, the results of the present study provide convincing evidence of a reduced luteal function during oestrous cycle and early pregnancy of sows with obesity/leptin resistance like Iberian sows, which may contribute to the low reproductive efficiency reported in this breed.     

Mycobacterium avium subsp. paratuberculosis infection in wildlife on three deer farms with a history of Johne's disease

Abstract

AIM: To determine the prevalence of Mycobacterium avium subsp. paratuberculosis (Map) infection in wildlife, in pastoral landscapes with a recent history of clinical Johne's disease in livestock.

METHODS: A total of 449 wild mammals and birds from three farms in the South Island of New Zealand with recent histories of clinical Johne's disease in their deer herds were trapped and examined for gross pathological changes in the gastrointestinal tract. Additionally, individual mesenteric lymph nodes from 380 mammals, and segments of gastrointestinal tract from 32 birds were excised, homogenised and cultured for viable Map bacilli. The prevalence of Map infection was then calculated for the various species. Faecal samples from those mammals which had culture-positive tissues were further cultured for the presence of Map.

RESULTS: Gross pathological changes were identified in the gastrointestinal tract of four brushtail possums, one cat, six ferrets, 12 hares, six hedgehogs, three rabbits, one stoat, and one paradise shelduck. Infection with Map in the gastrointestinal tract was confirmed in only three of these cases, one each of brushtail possums, hares and hedgehogs. In contrast, Map infection in the absence of gross pathological changes was frequently recorded in enteric tract tissues of mammals and birds. Among mammals, Map infection was recorded in 18/73 (25%) brushtail possums, 4/23 (17%) cats, 15/42 (36%) hedgehogs and 29/113 (26%) rabbits. Among birds, intestinal tract tissue Map infection was recorded in 3/17 (18%) paradise shelducks. Among 64 of the 74 mammals which had Map culture-positive tissues, 38% (n=5) of hedgehogs and 11% (n=3) of rabbits also had culture-positive faecal samples.

CONCLUSIONS: This study is the first to identify that Map infection can be prevalent in wildlife in New Zealand. There was a high prevalence of Map infection among both scavenging and grazing wild animals. Both mammals and birds are capable of harbouring viable Map organisms in their gastrointestinal tract; further, viable Map was excreted into the environment via faeces by hedgehogs and rabbits.

CLINICAL RELEVANCE: Previous studies overseas have postulated a role of wildlife as reservoirs of Map infection and possible vectors of Johne's disease to livestock. Here, brushtail possums, hedgehogs and rabbits and in particular were identified as potential wildlife hosts for Map infection in NewZealand. This suggests that several wildlife species could contribute to the persistence of Map infection within a wildlife/livestock complex, and potentially, perhaps more importantly, to the spread of infection between farms.     

Molecular characterization of genetic basis of Sugarcane Yellow Leaf Virus (SCYLV) resistance in Saccharum spp. hybrid

Sugarcane (Saccharum Spp.) produces 80% of the world's sugar along with other by‐products. The production of sugarcane is vulnerable to infestation of sugarcane yellow leaf virus (SCYLV) worldwide. A study was conducted using an F₁ segregating population derived from CP95‐1039 × CP88‐1762 to identify the genetic factors underlying SCYLV resistance. The disease infection data were measured using tissue blot immunoassay after 6 years of exposure to the virus under natural field conditions. Genetic maps were created using genotyping by sequencing‐based markers for each parent separately following a pseudo‐testcross approach. Two quantitative trait loci (QTL) were detected for SCYLV resistance accounting for 28% of the phenotypic variation. A major QTL qSCYLR79 located on linkage group 79 and linked with marker 3PAV3154 appears to be unique for SCYLV resistance in sugarcane. Progeny having a combination of two major alleles had 31% less SCYLV incidence than progeny with a combination of major and minor alleles in the genomic region of qSCYLR79. Thus, selection against the minor allele may decrease the SCYLV incidence in sugarcane.     

Candidatus Actinochlamydia pangasiae sp. nov. (Chlamydiales,Actinochlamydiaceae), a bacterium associated with epitheliocystis in Pangasianodon hypophthalmus

Chlamydial infections are recognised as causative agent of epitheliocystis, reported from over 90 fish species. In the present study, the farmed striped catfish Pangasianodon hypophthalmus (14–15 cm, 70–90 g) with a history of cumulative mortality of about 23% during June and July 2015, were brought to the laboratory. The histopathological examination of gills from the affected fish revealed presence of granular basophilic intracellular inclusions, mostly at the base of the interlamellar region and in gill filaments. A concurrent infection with Trichodina spp., Ichthyobodo spp. and Dactylogyrus spp. was observed in the gills. The presence of chlamydial DNA in the gills of affected fish was confirmed by amplification and sequencing of 16S rRNA gene. BLAST‐n analysis of these amplicons revealed maximum similarity (96%) with Candidatus Actinochlamydia clariae. On the basis of phylogenetic analysis, it was inferred that the epitheliocystis agents from striped catfish were novel and belonged to the taxon Ca. Actinochlamydia. It is proposed that epitheliocystis agents from striped catfish will be named as Ca. Actinochlamydia pangasiae. The 16S rRNA gene amplicons from novel chlamydiae were labelled and linked to inclusions by in situ hybridisation. This is the first report of epitheliocystis from India in a new fish host P. hypophthalmus.     

Effect of conjoint application of indigenous PGPR and chemical fertilizers on productivity of maize (Zea mays L.) under mid hills of Himachal Pradesh

The present study was conducted to work out the conjoint application of indigenous PGPR (plant growth promoting rhizobacteria) and chemical fertilizers levels on the productivity of maize (Zea mays L.). Three best PGPR isolates (B1N1, MAT1 and DHK) having maximum PGP (plant growth promoting) traits were screened at different recommended doses (80%, 60% and 40%) of NP (Nitrogen and Phosphorous) under net house conditions, and finally two isolates (B1N1 and MAT1) along with optimum dose i.e. 80% of NP were selected for field experimentation, which was performed for two years consecutively i.e. 2013–2015 under random block design (RBD). Conjoint application of 80% recommended doses of NP with PGPR (B1N1) significantly increased maize yield (11.7%), plant height (12.9%) and biomass (17.9%), over control (100% recommend dose of fertilizer (RDF) of chemical fertilizer). Therefore, the results revealed the potential of indigenous PGPR isolates to supplement about 20% NP fertilizers without hampering the productivity of maize.     

Ultrasound during mid‐gestation: Agreement with physical foetal and placental measurements and use in predicting gestational age in sheep

To determine the effects of poor maternal nutrition and litter size on foetal growth during mid‐gestation, pregnant ewes (n = 82) were fed 100%, 60% or 140% of NRC TDN beginning at day 30.2 ± 0.2 of gestation. Transabdominal ultrasound was performed weekly between day 46.0 ± 0.4 and 86.0 ± 0.7 to monitor foetal heart width (HW), umbilical diameter (UMB), rib width (RW) and placentome outer (OD) and inner diameter (ID). Data were analysed with repeated‐measures using the mixed procedure for effects of maternal diet, litter size and gestation, and equations predictive of gestational age were generated using the regression procedure. To determine the agreement of ultrasound measurement and actual size, ewes (n = 20–21) were euthanized at day 45 or 90 to obtain corresponding postmortem measurements for Bland–Altman analysis. The HW, UMB and placentome OD and ID increased with gestation (p < .0001) but were unaffected by maternal diet or litter size (p ≥ .12). Ultrasound underestimated postmortem measurements of HW (14.8%), UMB (7.3%), placentome OD (4.5%) and ID (37.3%) at day 90 of gestation. Ultrasound underestimated RW at day 45 (7.7%) but overestimated RW (23.8%) at day 90, indicating inconsistent bias when reporting RW by ultrasound. Combining the HW, UMB, RW and placentome OD generated the strongest equation predictive of gestational age (R² = .91). These findings indicate that during mid‐gestation, maternal diet or litter size did not affect HW, UMB or placentome diameters and these factors can be used to estimate gestational age.