Ectopic expression of an osmotin gene leads to enhanced salt tolerance in transgenic chilli pepper (<Emphasis Type="Italic">Capsicum annum</Emphasis> L.)

Plants, when exposed to abiotic or biotic stress, produce several pathogenesis-related proteins to counteract the effects of stress. Osmotin is one of the important pathogenesis-related proteins induced during several stress conditions. We have developed improved salt stress tolerant transgenic chilli pepper plants (Capsicum annum L. var. Aiswarya 2103) by ectopic expression of the Nicotiana tabaccum osmotin gene using Agrobacterium tumefaciens EHA105 as a vector. Four-week-old chilli pepper leaves were used as an explant and A. tumefaciens EHA105 harboring pBINASCOSM plasmid that contains osmotin gene under the control of CaMV 35S promoter and npt II as a selectable marker was used in co-cultivation. Transgene integration and expression were analyzed using molecular, immunochemical, and biochemical assays. PCR and Southern blot analysis confirmed that osmotin gene has been successfully integrated into the genome of chilli pepper plants. The osmotin gene was stably segregated and expressed in T₂ generation transgenic chilli pepper plants, and it was confirmed by Western blot analysis. Biochemical assays of these putative transgenic plants revealed enhanced levels of chlorophyll, proline, glycinebetaine, APX, SOD, DHAR, MDHAR, GR, and relative water content. Yield potential of the putative transgenic chilli pepper plants was evaluated under salinity stress conditions in a green house. The putative transgenic chilli pepper plants overexpressing the osmotin gene were morphologically similar to wild-type plants and produced 3.32 kg chilli pepper fruits per plant at 300 mM NaCl concentration.     

An Augmented SMS Intervention to Improve Access to Antenatal CD4 Testing and ART Initiation in HIV-Infected Pregnant Women: A Cluster Randomized Trial

BackgroundLess than one-third of HIV-infected pregnant women eligible for combination antiretroviral therapy (ART) globally initiate treatment prior to delivery, with lack of access to timely CD₄ results being a principal barrier. We evaluated the effectiveness of an SMS-based intervention to improve access to timely antenatal ART.MethodsWe conducted a stepped-wedge cluster randomized trial of a low-cost programmatic intervention in 20 antenatal clinics in Gaborone, Botswana. From July 2011-April 2012, 2 clinics were randomly selected every 4 weeks to receive an ongoing clinic-based educational intervention to improve CD₄ collection and to receive CD₄ results via an automated SMS platform with active patient tracing. CD₄ testing before 26 weeks gestation and ART initiation before 30 weeks gestation were assessed.ResultsThree-hundred-sixty-six ART-naïve women were included, 189 registering for antenatal care under Intervention and 177 under Usual Care periods. Of CD₄-eligible women, 100 (59.2%) women under Intervention and 79 (50.6%) women under Usual Care completed CD₄ phlebotomy before 26 weeks gestation, adjusted odds ratio (aOR, adjusted for time that a clinic initiated Intervention) 0.87 (95% confidence interval [CI]0.47–1.63, P = 0.67). The SMS-based platform reduced time to clinic receipt of CD₄ test result from median of 16 to 6 days (P<0.001), was appreciated by clinic staff, and was associated with reduced operational cost. However, rates of ART initiation remained low, with 56 (36.4%) women registering under Intervention versus 37 (24.2%) women under Usual Care initiating ART prior to 30 weeks gestation, aOR 1.06 (95%CI 0.53–2.13, P = 0.87).ConclusionsThe augmented SMS-based intervention delivered CD₄ results more rapidly and efficiently, and this type of SMS-based results delivery platform may be useful for a variety of tests and settings. However, the intervention did not appear to improve access to timely antenatal CD₄ testing or ART initiation, as obstacles other than CD₄ impeded ART initiation during pregnancy.     

Highly efficient Agrobacterium-mediated transformation of banana cv. Rasthali (AAB) via sonication and vacuum infiltration

A reproducible and efficient transformation method was developed for the banana cv. Rasthali (AAB) via Agrobacterium-mediated genetic transformation of suckers. Three-month-old banana suckers were used as explant and three Agrobacterium tumefaciens strains (EHA105, EHA101, and LBA4404) harboring the binary vector pCAMBIA1301 were used in the co-cultivation. The banana suckers were sonicated and vacuum infiltered with each of the three A. tumefaciens strains and co-cultivated in the medium containing different concentrations of acetosyringone for 3 days. The transformed shoots were selected in 30 mg/l hygromycin-containing selection medium and rooted in rooting medium containing 1 mg/l IBA and 30 mg/l hygromycin. The presence and integration of the hpt II and gus genes into the banana genome were confirmed by GUS histochemical assay, polymerase chain reaction, and southern hybridization. Among the different combinations tested, high transformation efficiency (39.4 ± 0.5% GUS positive shoots) was obtained when suckers were sonicated and vacuum infiltered for 6 min with A. tumefaciens EHA105 in presence of 50 μM acetosyringone followed by co-cultivation in 50 μM acetosyringone-containing medium for 3 days. These results suggest that an efficient Agrobacterium-mediated transformation protocol for stable integration of foreign genes into banana has been developed and that this transformation system could be useful for future studies on transferring economically important genes into banana.     

Resistance in accessions of cowpea to the coreid pod-bug Clavigralla tomentosicollis (Hemiptera: Coreidae)

Laboratory and screenhouse experiments were conducted to identify antibiosis and tolerance in four wild accessions of cowpea, Vigna unguiculata subsp. dekindtiana, to Clavigralla tomentosicollis St?l, the most damaging pod-sucking bug on cultivated Vigna genotypes in West Africa. The wild accession TVnu 151 showed antibiosis resistance, causing >50% mortality of the nymphs within 3 d of placing them on pods. Nymphs died more quickly on TVnu 151 than on TVnu 72, the wild and resistant control of the V. vexillata species which affected the weights of surviving insects to a much greater degree than TVnu 151. The three other accessions of the subspecies dekindtiana (TVnu 369, TVnu 517, and TVnu 707) did not cause significant mortality to the bugs, but rather extended their developmental time, with surviving adults showing lower weights and slower oviposition rates than those on the susceptible control IT84S-2246. Wild accessions affected male and female bugs differently, resulting in differential survival. This was reflected by the sex ratio which was male biased on the wild accessions (1:0.3-1:0.9), and female biased on IT84S-2246 (1:1.5). No evidence of tolerance was found in the four accessions of V. unguiculata subsp. dekindtiana. Overall, seed traits seemed to be the major resistance component in these wild accessions.     

Assessment of chemical resistance of wild and cultivated Vigna species to the brown pod-bug Clavigralla tomentosicollis Stal (Hem., Coreidae)

The effects of secondary metabolites in different Vigna species on the development of Clavigralla tomentosicollis were investigated in an artificial seed system using different fractions of crude pod extracts, while the orientation response of this pod‐bug to volatile extracts was studied using a dual‐choice olfactometer. Feeding on the neutral fraction extracts, in contrast to the basic and acidic fractions, resulted in significantly higher mortalities, longer total developmental time, and lower growth index of the insects in comparison with controls. All volatile extracts elicited an avoidance reaction by C. tomentosicollis, except the volatile from the susceptible genotype IT84S‐2246 which generally attracted as many insects as controls. Extracts from wild Vigna species showed higher activity than those from their cultivated relatives. The present study which has established that most secondary metabolites in cowpea pods were localized in the neutral fraction of the crude extract, could facilitate experiments on the separation and characterization of the toxic factors involved.     

Interaction between pod age and position on damage to cowpea Vigna unguiculata by hemipteran pod-sucking bugs

Laboratory and screenhouse studies were carried out to assess the relationship between pod age and pod position of cowpea and damage by different pod bug species. The coreids Clavigralla tomentosicollis St?l and Riptortus dentipes Fabricius caused significant damage to young pods of cultivated genotypes, in contrast to the coreid Anoplocnemis curvipes Fabricius and the pentatomid Aspavia armigera Fabricius which exhibited minor feeding activity. Percent seed damage declined with pod age, the critical stage for pod bug infestation being when pods were about eight days old. Clavigralla tomentosicollis and R. dentipes caused significantly higher damage to pods located within the leaf canopy, thus behaving differently from Anoplocnemis curvipes which showed a distinct preference for pods growing above the leaf canopy. The feeding activity of Aspavia armigera was not affected by the position of pods on the plant. Overall, the study suggests that cowpea genotypes with a short flowering period and pods held above the leaf canopy offer the most promise in the management of pod-sucking pests.     

Inferences on pod wall and seed defences against the brown cowpea coreid bug, Clavigralla tomentosicollis Stäl. (Hem., Coreidae) in wild and cultivated Vigna species

Studies were conducted in the laboratory to investigate the different components of the resistance of Vigna vexillata A. Richards, TVnu 72, and several cultivars of cultivated Vigna spp. to infestation and damage by the brown cowpea coreid, Clavigralla tomentosicollis Stäl. The main objective was to determine the different roles of the pod wall and seeds, and the relationship between seed size, number, and damage by this insect. Results showed that both the pod wall and seed clearly contribute different components to the resistance of TVnu 72 to C. tomentosicollis . Analysis of the results suggests that the seed defences and the pod wall pericarp may be more important than are trichomes in this resistance. Seed number was found to influence the extent of damage in a more predictable manner than seed size. However, because both of these traits are generally inversely related and mutually exclusive, their individual effects cannot be completely separated. The implications of these traits in a breeding programme targeted at a specific consumer group are discussed.     

Extracts from Tephrosia vogelii for the protection of stored legume seeds against damage by three bruchid species

Aliquots of powdered dried leaves of Tephrosia vogelii (Leguminosae – Papilionoideae) were extracted with hexane, acetone and ethanol. The extracts were tested for their potential to protect stored legume seeds from damage by the bruchids Acanthoscelides obtectus, Callosobruchus maculatus and Callosobruchus chinensis (Coleoptera – Bruchidae). While the acetone and ethanol extracts were ineffective except for a significant but low effect on A. obtectus, the hexane extract had a relatively high efficacy, providing similar control of all three bruchid species. This effect was manifested in slowly‐reduced adult survival within 7 days and in reduced numbers of eggs laid and of F₁ progeny produced, resulting in seed damage averaging 7.1% compared to 99.8% in the untreated controls after one generation. These findings are discussed in the context of previously identified constituents of T. vogelii, the rotenoids rotenone, tephrosin and deguelin.