High-frequency regeneration via multiple shoot induction of an elite recalcitrant cotton (Gossypium hirsutum L. cv Narashima) by using embryo apex

Cotton (Gossypium hirsutum L.) is one of the most commercially important fiber crops in the world. Compared with other crops, cotton represents a recalcitrant species for regeneration protocols. The development of efficient and rapid regeneration protocol for elite Indian cotton variety could help improve the quality characteristics and biotic or abiotic stress tolerance. Here we report a novel regeneration protocol in Indian cotton cultivar Narashima. The maximum number of multiple shoots obtained was 16 per explants, performance which has never been achieved in any prior reports. The embryo apex explants were isolated from 2 d old in vitro growing seedlings. Explants were cultured on MS medium containing different plant growth regulator combinations in order to induce multiple shoots. Among the tested combinations, the 2 mg/l of 6-benzylaminopurine (BAP) and 2 mg/l kinetin (KIN) proved to be most suited for achieving the maximum number of multiple shoots. The elongation of multiple shoots was obtained in media supplemented with gibberellic acid (GA3). The regenerated plants were successfully hardened in earthen pots after adequate acclimatization. This method avoids callus tissue, the stage of regeneration which may lead to somaclonal variation. The important feature of the presented method is shortening of regeneration time, as well as the induction of a high number of multiple shoots per explants. The present protocol may provide an efficient and rapid regeneration tool for obtaining more stable transformants from embryo apex explants of Indian cotton cultivar Narashima.     

<i>Aspergillus tubingensis</i> Causes Leaf Spot of Cotton (<i>Gossypium hirsutum</i> L.) in Pakistan

Cotton (Gossypium hirsutum L.) is a key fiber crop of great commercial importance. Numerous phytopathogens decimate crop production by causing various diseases. During July-August 2018, leaf spot symptoms were recurrently observed on cotton leaves in Rahim Yar Khan, Pakistan and adjacent areas. Infected leaf samples were collected and plated on potato dextrose agar (PDA) media. Causal agent of cotton leaf spot was isolated, characterized and identified as Aspergillus tubingensis based on morphological and microscopic observations. Conclusive identification of pathogen was done on the comparative molecular analysis of CaM and β-tubulin gene sequences. BLAST analysis of both sequenced genes showed 99% similarity with A. tubingensis. Koch’s postulates were followed to confirm the pathogenicity of the isolated fungus. Healthy plants were inoculated with fungus and similar disease symptoms were observed. Fungus was re-isolated and identified to be identical to the inoculated fungus. To our knowledge, this is the first report describing the involvement of A. tubingensis in causing leaf spot disease of cotton in Pakistan and around the world.     

Feeding behavior of Brevicoryne brassicae in resistant and susceptible collard greens genotypes: interactions among morphological and chemical factors

The cabbage aphid, Brevicoryne brassicae (L.) (Hemiptera: Aphididae), is distributed throughout the tropical and subtropical areas of the world. The main crops attacked by B. brassicae are cabbage, collard greens, broccoli, Brussels sprouts, and cauliflower. To survive the attack of pest insects, plants have evolved various resistance mechanisms that may affect pest feeding behavior. The use of electronic monitoring through EPG (electrical penetration graph) can help characterize and distinguish the resistance mechanisms involved. This study evaluated the feeding behavior of B. brassicae in eight genotypes of collard greens, Brassica oleraceae L. var. acephala (Brassicaceae), exhibiting antixenosis and/or antibiosis resistance to this insect. Possible correlations were established between the glucosinolate levels, the hardness, and the epicuticular wax on the leaves vs. aphid feeding behavior. On the genotypes 22V, 5E, and 27VA, for which many ‘potential drop’ waves were performed, aphid development was slower, indicating antixenosis as resistance type. Aphids on the genotypes 22V and 24X required more time until accessing the phloem, also suggesting antixenosis as resistance category. Genotypes 22V and PE had hard leaves, which also points at antixenosis. Genotypes 20T and HS had higher total wax and wax mg⁻¹. Feeding parameters on ARI and 24X were similar to those observed on HS; antibiosis is likely to be the predominant resistance category of this germplasm. Because HS was considered as a susceptible standard genotype in this study, a higher gluconapin amount indicates that this compound does not influence cabbage aphid feeding behavior. The present study confirms that analysis of the physical and chemical aspects of collard greens genotypes by the EPG technique can provide a useful approach for the study of plant resistance to cabbage aphids.     

Distributions of western flower thrips (Thysanoptera: Thripidae) and its predatory bug Orius niger (Hemiptera: Anthocoridae) assessed by coloured sticky traps and plant samplings in cotton

The capturing efficiency of four coloured (yellow, green, white and blue) sticky traps, placed at the top, middle and bottom strata of cotton plants, was tested for the western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), and its predatory bug, Orius niger (Wolff) (Hemiptera: Anthocoridae), as well as spatial distributions of both insects on the plant in years 2006 and 2007. The white coloured trap was the most attractive to F. occidentalis and O. niger in the 2-year study. The blue coloured trap was the least attractive for Orius. The mean numbers of F. occidentalis and O. niger on the top plant parts (flowers and leaves) and in all coloured traps positioned on the upper parts of the plants were greater than those found in the lower two strata. Taylor's power law analysis showed that F. occidentalis and O. niger adults aggregated in the flowers or on the leaves. This study suggests that top flowers could be preferentially sampled to determine population densities of Frankliniella flower thrips and Orius species in cotton, and thus, sticky traps should be placed on the top level of plants. F. occidentalis: O. niger ratios in the flowers varied from 4 to 60 thrips per Orius adult in the three plant strata. This result may indicate that F. occidentalis experiences more predation from Orius.     

Secretion of Spodoptera littoralis female reproductive system on deposited egg masses that possibly acts as oviposition deterring substance for conspecific females

Egg masses laid by Spodoptera littoralis mated female moths were extracted by petroleum ether (PE), ethanol (E) and Ringer's solution (RS). Egg-wash extracts were evaporated and the weights of crude materials were obtained. Different aqueous concentrations were made. The amount of extracted material increased as the weight of eggs used increased and vice versa. Coating Nerium oleander leaves with aqueous egg-wash extracts prepared from S. littoralis egg-masses deterred the mated conspecific female moths from ovipositing their eggs on treated leaves, as well as causing a decrease in the total number of deposited eggs per female during the moth's life span. The highest deterrent effect on conspecific female moths to oviposit their eggs was obtained after treatment of N. oleander leaves with PE or E egg-wash extract. The deterrent effects of the tested egg-wash extracts was concentration dependant; an increase in the concentration of any extract caused an obvious decrease in the number of deposited egg-masses and the total number of laid eggs on the treated N. oleander leaves.     

Laboratory studies on extracts of Melia fruit and Neem fruit on B. brassicae,B. tabaci and their predators and parasites

Abstract

The biological effect of plant extracts on the two predators C. carnea and C. undecimpunctata was studied in the laboratory. Treated B. brassicae was offered to the newly hatched larvae of the two predators. The rate of prey consumption and duration of the larval instars of the two predators were slightly affected as a result of the treatments of the plant extracts. Adults of the aphid parasite D. Rapae and the whitefly parasite E. Mundus were exposed to the plant extracts via feeding on treated honey droplets or parasitized treated hosts. Mortality percentages of D. Rapae fed on honey contaminated with plant extracts were 50 and 80% for extracts of Melia fruits in ethanol and in petroleum ether, respectively. Mortalities in parasites developed in hosts treated with plant extracts of Neem fruits in ethanol and Melia flowers in petroleum ether in were 33.3 and 81.5%, respectively. Similar results were obtained when E. Mundus received the same treatments of the plant extracts.     

Plasticity in biochemical characteristics of cotton aphid,Aphis gossypii Glover in response to different host plants and neonicotinoid pesticides

In the present investigation an effort was made to realise the role of feeding host plants on some enzymes’ activities. The results showed that the enzymes’ activities were changed in the aphids feeding on different host plants which assist in detoxification of their host metabolites. It is important when the aphids are exposed to insecticides. The results indicated that the measured enzyme activity has significant changes depending on the host plant. It is shown in this study that there are no significant differences between different host plants on esterase activity (p = 0.446); however, there is a significant difference between GSH activity (p = 0.047) but this relationship is not significant on MFO activity (p = 0.417). Among three strains of Ag-PP, Ag-MO and Ag-FA, strain Ag-PP was the most resistant strain against neonicotinoids, and the resistance mechanism was related to metabolic increase in carboxyl esterase activity. The results showed that strain of Ag-MO was the most susceptible strain against neonicotinoids. The result of this investigation also showed that the general esterases might play an important role in conferring or contributing to neonicotinoid resistance in the cotton aphids.     

Races of Xanthomonas citri subsp. malvacearum,the causal organism of bacterial blight of cotton in northern Nigeria

Xanthomonas citri subsp. malvacearum (Xcm) causes severe qualitative and quantitative losses to farmers in cotton-growing areas of the world. Isolates of Xcm were extracted from cotton seeds obtained from five ginneries located in Funtua, Malumfashi, Gusau and Zaria and standardised to 10^?5?cfu/ml. One isolate per location was used to inoculate three sets of 10 cotton differential lines known to differentiate races of Xcm through possession of B-genes for resistance to bacterial blight. Each cotton differential line was inoculated with the isolates at the six-leaf stage and SDW was inoculated as control. One hundred and sixty pots used were arranged in completely randomised design on the screen house bench. Four different pathogenic races were identified namely race 1, race 12, race 13 and race 16. This confirms the existence of an evolution of the species across northern Nigeria.     

Potentialities of methoxyfenozide for the integrated management of Helicoverpa armigera (Lepidoptera: Noctuidae) on cotton in Benin,West Africa

Intensive use of chemical insecticides against Helicoverpa armigera has led to the development of resistance to the major chemical families of insecticides. Consequently, management of H. armigera using conventional chemical insecticides is increasingly difficult. Methoxyfenozide is an agonist of the insect moulting hormone 20-hydroxyecdysone that acts faster than chitin synthesis inhibitors. The present work is aimed to assess the effectiveness of methoxyfenozide for use against H. armigera on cotton in Benin, West Africa. Laboratory tests and field experiments have been carried out. For laboratory studies, topical application of methoxyfenozide was done using L2 and L5 instar larva of H. armigera. Tested methoxyfenozide concentrations varied from 24 μl/ml to 144 μl/ml and the control is water only. For field experiments, a complete randomised block design was used with methoxyfenozide (72 μl/ml) and a control (no spraying). Berthoud Ultra Low Volume sprayer was used to spray methoxyfenozide suspension at 60-l per hectare. Two sprays were applied, 7 days apart. Laboratory tests indicated that 24 h after application of methoxyfenozide, about 100% of treated L2 and L5 larva were morbid or dead compared with 0% for the control larva. There are no significant differences between tested concentrations. Morbid larvae died within 2 days. In field experiments, cotton yield harvested on methoxyfenozide treated plants was double of that obtained on untreated plants (9375 kg and 4875 kg per hectare respectively). Thus methoxyfenozide may be used as component of Integrated Pest Control Programme on cotton in Benin, West Africa.