Potential of Cellulase From Bacillus megaterium for Hydrolysis of Sargassum

Bacterial cellulase can be a promising biotechnological approach in seaweed degradation to facilitate extraction of bioactive molecules. In the present study, bacterial cultures were isolated from marine and terrestrial samples and screened for cellulase production. The selected cultures were grouped by random amplification of polymorphic DNA (RAPD) profiling, and the cultures showing a distinct banding pattern were identified by 16S rDNA sequence analysis. Cellulase from Bacillus megaterium (CB-sw1-I), a native isolate, was purified by ion-exchange chromatography with a purification fold of 6.91 and specific activity of 1.66 U. The purified enzyme with molecular weight of 43 kDa was optimally active at pH 6.0 and temperature of 60°C. The potentiality of enzyme in degradation of Sargassum thallus was evident with an increase in reducing sugar. Morphology of the thallus after 10 days digestion observed by scanning electron microscope also provided additional evidence for degrading ability of the enzyme. This enzyme could subsequently be used in hydrolysis of seaweed for extraction of bioactive molecules.     

Progress Towards Genetics and Breeding for Minor Genes Based Resistance to Ug99 and Other Rusts in CIMMYT High-Yielding Spring Wheat

Wheat rusts continue to cause significant losses worldwide despite major efforts given to their genetic control. This is due to frequent evolution and selection of virulence in pathogen overcoming the deployed race-specific resistance genes. Although the life of effective race-specific resistance genes can be prolonged by using gene combinations, an alternative approach being implemented at CIMMYT is to deploy varieties that posses adult plant resistance （APR） based on combinations of minor, slow rusting genes. When present alone, the APR genes do not confer adequate resistance especially under high disease pressure; however, combinations of 4 or 5 minor genes usually result in ＂near-immunity＂ or a high level of resistance. Although only a few APR genes are catalogued, various APR QTLs are now known and could lead to further characterization of additional genes. Four characterized genes have pleiotropic effects in conferring partial APR to all 3 rusts and powdery mildew, thus simplifying the task of breeding wheat varieties that are resistant to multiple diseases. Significant progress was made recently in developing high-yielding wheat germplasm that possesses high levels of APR to all three rusts by implementing a Mexico- Kenya shuttle breeding scheme. Parents with APR to Ug99 were hybridized with high-yielding parents that had adequate to high levels of APR to leaf rust and yellow rust. Segregating populations and advanced lines from these crosses were selected under high rust pressures in Mexico （leaf rust and yellow rust） and Kenya （Ug99 stem rust and yellow rust） to identify high- yielding progenies that possess high to adequate APR to all three rusts. International distribution of these high-yielding wheats is underway through CIMMYT intemational yield trials and screening nurseries. It is expected that several wheat varieties with APR to three rusts will be released and grown in various countries in the near-future that will allow determining the durability of resistance.     

Fabrication and in-pond operation of a low-cost,semiautomated portable carp hatchery with a temperature-adjustability feature

The present study describes a simple, portable, and inexpensive in-pond hatchery model assembled from locally available materials for seed production of small and medium-sized carp at high efficiency. This light-weight hatchery requires minimal engineering skills for assembly and operation. Further, the hatchery water supply mechanism has built-in components to filter out sediments and zooplankton from the pond water besides maintaining the water temperature 5°C–7°C lower than the pond surface water by drawing water from a metalimnion layer without additional energy requirements. Spontaneous spawning, automatic transfer of eggs to the incubation tank, and 80% hatching rates at an egg-holding capacity of 2,000 eggs/L was recorded for Barbonymus gonionotus, Labeo bata, and Labeo rohita with minimal human intervention. The production capacity of 0.06 million fry per cycle and estimated benefit-cost ratio of 2.48 demonstrated the high efficiency and economic feasibility of this portable hatchery.     

Validation and Marker-Assisted Selection of Stem Rust Resistance Gene Sr2 in Indian Wheat Using Gel-Based and Gel-Free Methods

Journal of Crop Science and Biotechnology - Stem rust resistance gene Sr2 is an important slow-rusting gene which has provided resistance against stem rust for many years. The Sr2 gene has durable...     

Genetic analysis of prolificacy in “Sikkim Primitive”—A prolific maize (Zea mays) landrace of North‐Eastern Himalaya

Prolificacy assumes significance for development of high‐yielding baby corn hybrids. “Sikkim Primitive” is a native landrace of North‐Eastern Himalaya, and is the highest prolific maize germplasm. So far, the genetics of prolificacy in “Sikkim Primitive” has not been deciphered. Here, a prolific inbred (MGU‐SP‐101) developed from “Sikkim Primitive” was crossed with four non‐prolific inbreds viz., LM13, BML7, HKI161 and HKI1128. Six generations (P₁, P₂, F₁, F₂, BC₁P₁ and BC₁P₂) of the crosses were evaluated at two locations during rainy season 2018. MGU‐SP‐101 possessed 2.50–3.78 ears per plant compared to 1.06–1.86 among non‐prolific inbreds. The variation for ears was the highest in F₂s (1–8), followed by BC₁P₁ (1–7) and BC₁P₂ (1–5). The quantitative inheritance pattern of prolificacy with prevalence of non‐allelic interactions of duplicate epistasis type has been observed. Dominance × dominance effect was predominant over additive × additive and additive × dominance effects. Total number of major gene blocks ranged from 0.41 to 2.86, thereby suggesting the involvement of at least one major gene/QTL governing the prolificacy. This is the first report of genetic dissection of prolificacy in “Sikkim Primitive”.     

Limited contribution of water availability in genotype-by-environment interaction in sugarcane yield and yield components

Water deficit is an important abiotic stress factor limiting sugarcane production. Understanding the effect of water deficit on cane yield, yield components and genotype × environment (G × E) interaction can aid in selecting genotypes that are adapted to water deficit conditions. In this study, response of cane yield and yield components of 10 sugarcane genotypes grown under 3 irrigation treatments—fully irrigated, semi-irrigated and rainfed—was examined across 3 locations and 3 crop-years within each location. Cane yield reduced by 32% and 9% in the rainfed and semi-irrigated treatments, respectively, compared to the fully irrigated treatment. Genotypic variation was highly significant for cane yield and yield components in all three irrigation regimes. Cane yield in the rainfed treatment showed significant genetic correlation with stalk diameter (r_g = 0.68), leaf number (r_g = 0.64), node number (r_g = 0.67) and stalk number (r_g = 0.68). Genotype × irrigation variance was negligible compared to genotype, genotype × location and genotype × crop-year variances. Commercial genotypes had higher yield in most environments except in low-yield potential environments, which caused a significant genotype × location × irrigation effect, suggesting opportunities for improving sugarcane productivity in these environments.     

Unveilingsources of stripe rust resistance in diverse wheat (<Emphasis Type="Italic">Triticum Aestivum</Emphasis> L.) germplasm using narrow down methodology: a proof of concept

Stripe rust of wheat caused by the fungal pathogen is a destructive foliar disease of wheat. Thus, it is crucial step to characterize the resistant germplasm for stripe rust in a diverse germplasm pool for their ultimate utilization in efficient crop rust resistance breeding. In the present study, we followed two pronged strategies involving integrated phenotypic and molecular characterization of 440 diverse wheat germplasm lines for rust resistance. The germplasm panel was extensively evaluated in field epiphytotic conditions during two consecutive years. After rigorous screening, 72 accessions were successfully revealed as resistant to moderately resistant to stripe rust. Subsequently, entries were then evaluated for their field agronomicperformances, considering prerequisites for serving as a donor germplasm,particularly for yield and 33 potential rust-resistant accessions were identified. Furthermore, to detect the sources of resistance, accessions were molecular characterized for potential race-specific resistance genes Yr5, Yr10,Yr15, and effective adult plant resistance (APR) gene Lr34/Yr18/pm38. We identified the 22 accessions possessing one or more single resistance genes and two accessions were observed with at least three of them. Moreover, Lr34/Yr18/pm38 was determined to confer resistance when observed along with any of the race-specific genes. Thus, the study not only provides proof of concept methodology to identify candidate resistant sources from large germplasm collections but simultaneouslyconfirmed the contribution of combining race-specific andnon-specific APR genes. The finding could further assist in the potential deployment of resistant genes directly into the stripe rust breeding program by involving marker-assisted approaches.     

Development of Bt transgenic potatoes for effective control of potato tuber moth by using cry1Ab gene regulated by GBSS promoter

Transgenic of Indian potato cultivar Kufri Badshah expressing synthetic, modified cry1Ab gene were developed against potato tuber moth (Phthorimaea opercullela Z.) a destructive pest. The cry1Ab gene was in spatial and temporal expression under the control of tuber-specific GBSSi promoter. The transformation vector pBinCG1 was developed harbouring transgene expression cassette comprising cry1Ab gene under the control of potato GBSSi promoter, castor bean catalase intron (5′UTR) and OCS termination signals. Selected kanamycin-resistant putative transformed potato lines were evaluated by positive PCR amplification of 713 bp, 1206 bp and 700 bp with npt II, GBSSi promoter and cry1Ab gene specific primers, respectively. Gene integration in transformed potato plants was elucidated through Southern hybridization and in planta transgene expression analysis. Transgene expression was highest in stolons, followed by tubers matured leaves and young leaves. Insect bioassay of stored transgenic tubers resulted in significant retardation and mortality in neonate tuber moth larvae. RT-PCR and northern hybridization confirmed stable expression in stored tubers. These transgenic lines; thus can represent an effective resistance management strategy which can significantly reduce the selection pressure on target and non-target insect pests to a threshold level.     

Changes in endogenous estrogens and expression of behaviors associated with estrus during the periovulatory period in Heatsynch treated Murrah buffaloes (Bubalus bubalis)

The objective of this study was (1) to establish the duration of behavioral estrus signs and timing of ovulation in Murrah buffaloes (n = 10) and (2) to determine relationship between behavioral estrus signs with change in plasma estrogen concentrations in animals treated with Heatsynch protocol. Estrus and its behavioral signs were detected at hourly intervals by visual observations per rectal examination of genitalia and bull parading four times in a day for 30 min each. Among the behavioral signs of estrus, swollen vulva was the best indicator of estrus followed by excitement and chasing by bull (90%). Among the duration of behavioral estrus signs, the first and longest duration of estrus signs was swollen vulva, which was seen up to 21.6 ± 1.1 h after onset of estrus. The mean total duration of estrus symptoms from appearance to disappearance of all the behavioral estrus symptoms was 40.2 ± 1.8 h. Endocrine profile during the periestrus period showed that the mean peak concentrations of total estrogen 330.9 ± 108.3 pg/ml occurred at 9.6 ± 1.0 h after estradiol benzoate injection. The average number of estrus symptoms observed per animal during Heatsynch treatment was 7.1. Ovulation occurred after 50.0 ± 2.0 h after estradiol benzoate treatment and 26.7 ± 2.0 h after end of total estrogen surge, respectively. In conclusion, our results suggest that all signs of behavioral estrus occurred after the preovulatory rise in estrogens. The first sign of estrus was swollen vulva, and this symptom persisted the longest.     

Genetic and Biological Diversity of Trichoderma stromaticum, a Mycoparasite of the Cacao Witches'-Broom Pathogen

ABSTRACT The witches'-broom disease, caused by the basidiomycete Crinipellis perniciosa, is the most limiting factor for cacao cultivation in Brazil. Trichoderma stromaticum is a mycoparasite of the witches'-broom pathogen of cacao that is currently being applied in the field to manage the disease in Bahia State, Brazil. In this work, molecular and traditional methods were used to study the genetic and biological diversity of this mycoparasite. Ninety-one isolates, mostly collected from farms not sprayed with the fungus, were analyzed by amplified fragment length polymorphisms (AFLP), which showed that two genetic groups (I and II) of T. stromaticum occur in Bahia State. This classification of T. stromaticum into two distinct AFLP groups was also in agreement with several other characteristics, including growth on agar media at different temperatures and sporulation on infected stem segments (broom pieces) and rice grains. Group II favors higher temperatures compared with group I. The genetic and biological differences of the isolates, however, were not evident in field experiments, where sporulation was evaluated on the surface of brooms under natural conditions. Our results show that there is considerable genetic and biological diversity within T. stromaticum in Bahia and other cacao-growing regions of South America that are affected by the witches'-broom disease. This diversity could be explored in the development of efficient biological control agents against the disease. Factors that may affect the application and performance of this biocontrol agent in the field, such as sporulation on rice substrate and on the brooms and growth at various temperatures, are discussed.