Banana weevil (Cosmopolites sordidus) is the most devastating pest of banana and plantain worldwide, yet current control measures are neither effective, sustainable, nor environmentally sound, and no resistant farmer-preferred cultivars are known to date. In this paper, we examined the ability to induce RNA interference (RNAi) in the banana weevil via feeding. We first developed an agar- and banana corm (rhizome) flour-based artificial diet in a multi-well plate setup that allowed the banana weevils to complete their life cycle from egg through the larval instars to the pupal stage in an average period of 53 days. Adults emerged about 20 days later. The artificial diet allowed the tunneling and burrowing habits of the larvae and successful metamorphosis up to adult eclosion. Adding dsRNA for laccase2 to the artificial diet resulted in albino phenotypes, confirming gene-silencing. Finally, C. sordidus was fed with dsRNA against a selection of essential target genes: snf7, rps13, mad1, vha-a, vha-d, and lgl for a period of 45 days. 100% mortality within 9–16 days was realized with dssnf7, dsrps13, and dsmad1 at 200 ng/mL artificial diet, and this corresponded to a strong reduction in gene expression. Feeding the dsRNA targeting the two vha genes resulted in 100% mortality after about 3–4 weeks, while treatment with dslgl resulted in no mortality above the dsgfp-control and the water-control. Our results have implications for the development of RNAi approaches for managing important crop pests, in that banana weevils can be controlled based on the silencing of essential target genes as snf7, rps13, and mad1. They also highlight the need for research into the development of RNAi for banana protection, eventually the engineering of host-induced gene-silencing (HIGS) cultivars, given the high RNAi efficacy and its species-specific mode of action, adding the RNAi approach to the armory of integrated pest management (IPM).
Low female fertility in bananas is the biggest hurdle for banana breeding. The aim of this study was to determine seed set patterns in East African Highland Cooking bananas (EAHBs) to inform future decisions on a more targeted approach of increasing seed set and subsequently banana-breeding efficiency. Matooke (AAA) and Mchare (AA) bananas are genetically distinct but belong to the same genetic complex, referred to as EAHBs. Seed set patterns in “Enzirabahima” (AAA), “Mshale” (AA), and “Nshonowa” (AA), all with residual fertility, were examined after hand pollination with a highly male fertile wild banana “Calcutta 4” (AA). Seed set in “Enzirabahima” is predominant in distal hands. Mchare cultivars have a slightly more even distribution of seeds in their hands compared to “Enzirabahima”. There is a gradual increase in seed set from proximal to distal hands with a slight drop in the last hand. This pattern is more definite in “Enzirabahima” and “Mshale”, while “Nshonowa” has a somewhat inconsistent pattern. There is also a drop in seed set per 100 fruits per hand from small to larger bunches. However, larger bunches have a higher pollination success compared to smaller bunches. They therefore set more seed on 100 fruits per hand and per bunch basis, if bunches without seed are accounted for. Pollination success rate increases from smaller to larger bunches of EAHBs. Seed set is biased toward the distal third part of fruits of examined EAHBs, as well as tetraploid Matooke hybrid “401K-1” (AAAA), and improved diploid “Zebrina” GF (AA) that were used for comparison. In comparison, in the highly female fertile “Calcutta 4”, seed set is along the entire length of the fruit. Seed set bias in the distal hands and distal end of fruits suggests a systematic mechanism rather than a random occurrence. It is expected that this information will provide a foundation for increased crossbreeding efficiency in bananas.
The burrowing nematode Radopholus similis is considered a major problem of intensive banana cultivation. It can cause extensive root damage resulting in the toppling disease of banana, which means that plants fall to the ground. Soaking R. similis in double-stranded (ds) RNA of the nematode genes Rps13, chitin synthase (Chs-2), Unc-87, Pat-10 or beta-1,4-endoglucanase (Eng1a) suppressed reproduction on carrot discs, from 2.8-fold (Chs-2) to 7-fold (Rps13). The East African Highland Banana cultivar Nakitembe was then transformed with constructs for expression of dsRNA against the same genes, and for each construct, 30 independent transformants were tested with nematode infection. Four months after transfer from in vitro culture to the greenhouse, the banana plants were transferred to a screenhouse and inoculated with 2000 nematodes per plant, and thirteen weeks later, they were analyzed for several parameters including plant growth, root necrosis and final nematode population. Plants with dsRNA constructs against the nematode genes were on average showing lower nematode multiplication and root damage than the nontransformed controls or the banana plants expressing dsRNA against the nonendogenous gene. In conclusion, RNAi seems to efficiently protect banana against damage caused by R. similis, opening perspectives to control this pest.
Banana Xanthomonas wilt (BXW) is a destructive disease caused by Xanthomonas vasicola pv. musacearum (Xvm) bacterium that indiscriminately infects all banana varieties grown in East and Central Afr...
Seed set in East African Highland Cooking bananas (EAHBs) is extremely low and therefore hampers breeding. Pollen–pistil interaction is a key contributing factor. We assessed the effect of pollen germination media (PGM) on seed set in EAHBs. Five EAHB cultivars were pollinated with pollen from the wild banana ‘Calcutta 4’. Glucose-based PGM sprayed on freshly emerged stigmas significantly increased seed set per 100 fruits per bunch. Increases were 73.5% in ‘Enzirabahima’, 39.9% in ‘Mshale’, and 302.4% in ‘Nshonowa’. However, PGM did not increase seed set in the female sterile ‘Mlelembo’ and ‘Nakitembe’. As larger bunches were more fertile, good field management practices are also recommended to get more seed to improve breeding efficiency.
Seed set in banana is influenced by weather, yet the key weather attributes and the critical period of influence are unknown. We therefore investigated the influence of weather during floral development for a better perspective of seed set increase. Three East African highland cooking bananas (EAHBs) were pollinated with pollen fertile wild banana ‘Calcutta 4′. At full maturity, bunches were harvested, ripened, and seeds extracted from fruit pulp. Pearson’s correlation analysis was then conducted between seed set per 100 fruits per bunch and weather attributes at 15-day intervals from 105 days before pollination (DBP) to 120 days after pollination (DAP). Seed set was positively correlated with average temperature (P < 0.05–P < 0.001, r = 0.196–0.487) and negatively correlated with relative humidity (RH) (P < 0.05–P < 0.001, r = −0.158–−0.438) between 75 DBP and the time of pollination. After pollination, average temperature was negatively correlated with seed set in ‘Mshale’ and ‘Nshonowa’ from 45 to 120 DAP (P < 0.05–P < 0.001, r = −0.213–−0.340). Correlation coefficients were highest at 15 DBP for ‘Mshale’ and ‘Nshonowa’, whereas for ‘Enzirabahima’, the highest were at the time of pollination. Maximum temperature as revealed by principal component analysis at the time of pollination should be the main focus for seed set increase.