Isolated and combined effects of soil salinity and waterlogging in seedlings of 'Green Dwarf' coconut / Efeitos isolados e combinados da salinidade do solo e encharcamento em mudas de coqueiro 'Anão Verde'

Soil salinization is a problem commonly found in semi-arid regions. In addition, the problem of salinity is aggravated in clayey soils when accompanied by cycles of waterlogging in the rainy season or when excess irrigation is applied. In this work we evaluated the isolated and combined effects of soil salinity and waterlogging on the responses of young plants of ‘Green Dwarf coconut. The experiment was conducted under controlled environment in a complete randomized block design, arranged in split plots with five replications. The plots comprised five waterlogging cycles (0, 1, 2, 3 and 4), each with a duration of four days, and applied at 30, 60, 90 and 120 days into the experimental period, with the sub-plots consisting of five levels of soil salinity (1.70, 11.07, 16.44, 22.14 and 25.20 dS m-1). Response of coconut seedlings to waterlogging was dependent on the level of soil salinity, with waterlogging significantly impairing biomass accumulation and leaf expansion at low soil salinity levels, but causing no additional harm at elevated salinity. Leaf gas exchange was reduced mainly due to soil salinity, and this response was related to stomatal and non-stomatal effects. Seedlings of ‘Green Dwarf coconut used in this study were classified as moderately-tolerant to salinity when grown in soils with an electrical conductivity up to 11.07 dS m-1, having the potential to be used in revegetation programs of salt-affected areas, provided that these areas are not exposed to frequent waterlogging cycles.(AU)

A salinização dos solos é um problema comumente encontrado em regiões semiáridas. Além disso, nos solos mais argilosos, o problema da salinidade vem acompanhado de ciclos de encharcamento do solo, no período de chuvas ou no caso de irrigação excessiva. Neste trabalho, avaliamos os efeitos da salinidade do solo e do encharcamento, de maneira isolada e combinada, nas respostas adaptativas de plantas jovens de coqueiro-anão-verde. O experimento foi conduzido em ambiente protegido, sob delineamento estatístico de blocos casualizados, arranjados em parcelas subdivididas com cinco repetições. As parcelas foram constituídas por cinco ciclos de encharcamento (0, 1, 2, 3 e 4), com duração de quatro dias cada, aos 30, 60, 90 e 120 dias do período experimental e as subparcelas foram constituídas por cinco níveis de salinidade do solo (1,70; 11,07; 16,44; 22,14 e 25,20 dS m-1). As respostas das mudas de coqueiro ao encharcamento dependeram do nível de salinidade do solo, os quais reduziram significativamente o acúmulo de biomassa e a expansão foliar em baixos níveis de salinidade do solo. Contudo, os níveis de encharcamento não causaram danos adicionais sob elevados níveis de salinidade. As trocas gasosas foliares foram reduzidas principalmente devido à salinidade do solo, e esta resposta pode estar relacionada aos efeitos estomáticos e não estomáticos. As mudas de coqueiro ‘Anão Verde utilizadas neste experiment foram classificadas como moderadamente tolerantes à salinidade, quando cultivadas em solos com condutividade elétrica de até 11,07 dS m-1, podendo ser utilizadas em programas de revegetação de áreas salinizadas, desde que essas áreas não estejam expostas a frequentes ciclos de encharcamento.(AU)

Protocol for the Micropropagation of Coconut from Plumule Explants.

Coconut is a crop that is economically important in several countries throughout the world. Unfortunately, production is decreasing because palms are affected by very serious phytoplasma diseases, such as lethal yellowing, and also most of coconuts are already very old. On the other hand, markets for coconut products have been rapidly growing in recent years. Hence, replanting of most cultivation surface worldwide, as well as establishing new surface, is urgently needed. This is an immense task, requiring at least a billion coconut palms that cannot be accomplished by traditional propagation through seed. Therefore the biotechnological alternative of micropropagation by somatic embryogenesis is needed. Research has been carried out on this subject in laboratories in several countries studying different approaches, testing different types of explants. The most responsive tissue has been plumule from zygotic embryos. A protocol for micropropagation of coconut based on plumule explants is described here. It involves the use of different media that are based on Y3 medium complemented with activated charcoal, gelling agent, sucrose, and growth regulators. These media allow the formation of embryogenic callus and somatic embryos, growth of shoots, and development of plantlets.

Isolated and combined effects of soil salinity and waterlogging in seedlings of 'Green Dwarf' coconut / Efeitos isolados e combinados da salinidade do solo e encharcamento em mudas de coqueiro 'Anão Verde'

Soil salinization is a problem commonly found in semi-arid regions. In addition, the problem of salinity is aggravated in clayey soils when accompanied by cycles of waterlogging in the rainy season or when excess irrigation is applied. In this work we evaluated the isolated and combined effects of soil salinity and waterlogging on the responses of young plants of ‘Green Dwarf’ coconut. The experiment was conducted under controlled environment in a complete randomized block design, arranged in split plots with five replications. The plots comprised five waterlogging cycles (0, 1, 2, 3 and 4), each with a duration of four days, and applied at 30, 60, 90 and 120 days into the experimental period, with the sub-plots consisting of five levels of soil salinity (1.70, 11.07, 16.44, 22.14 and 25.20 dS m-1). Response of coconut seedlings to waterlogging was dependent on the level of soil salinity, with waterlogging significantly impairing biomass accumulation and leaf expansion at low soil salinity levels, but causing no additional harm at elevated salinity. Leaf gas exchange was reduced mainly due to soil salinity, and this response was related to stomatal and non-stomatal effects. Seedlings of ‘Green Dwarf’ coconut used in this study were classified as moderately-tolerant to salinity when grown in soils with an electrical conductivity up to 11.07 dS m-1, having the potential to be used in revegetation programs of salt-affected areas, provided that these areas are not exposed to frequent waterlogging cycles.

A salinização dos solos é um problema comumente encontrado em regiões semiáridas. Além disso, nos solos mais argilosos, o problema da salinidade vem acompanhado de ciclos de encharcamento do solo, no período de chuvas ou no caso de irrigação excessiva. Neste trabalho, avaliamos os efeitos da salinidade do solo e do encharcamento, de maneira isolada e combinada, nas respostas adaptativas de plantas jovens de coqueiro-anão-verde. O experimento foi conduzido em ambiente protegido, sob delineamento estatístico de blocos casualizados, arranjados em parcelas subdivididas com cinco repetições. As parcelas foram constituídas por cinco ciclos de encharcamento (0, 1, 2, 3 e 4), com duração de quatro dias cada, aos 30, 60, 90 e 120 dias do período experimental e as subparcelas foram constituídas por cinco níveis de salinidade do solo (1,70; 11,07; 16,44; 22,14 e 25,20 dS m-1). As respostas das mudas de coqueiro ao encharcamento dependeram do nível de salinidade do solo, os quais reduziram significativamente o acúmulo de biomassa e a expansão foliar em baixos níveis de salinidade do solo. Contudo, os níveis de encharcamento não causaram danos adicionais sob elevados níveis de salinidade. As trocas gasosas foliares foram reduzidas principalmente devido à salinidade do solo, e esta resposta pode estar relacionada aos efeitos estomáticos e não estomáticos. As mudas de coqueiro ‘Anão Verde’ utilizadas neste experiment foram classificadas como moderadamente tolerantes à salinidade, quando cultivadas em solos com condutividade elétrica de até 11,07 dS m-1, podendo ser utilizadas em programas de revegetação de áreas salinizadas, desde que essas áreas não estejam expostas a frequentes ciclos de encharcamento.

Predator performance is impaired by the presence of a second prey species.

The simultaneous infestation of a plant by several species of herbivores may affect the attractiveness of plants to the natural enemies of one of the herbivores. We studied the effect of coconut fruits infested by the pests Aceria guerreronis and Steneotarsonemus concavuscutum, which are generally found together under the coconut perianth. The predatory mite Neoseiulus baraki produced lower numbers of offspring on fruits infested with S. concavuscutum and on fruits infested with both prey than on fruits with A. guerreronis only. The predators were attracted by odours emanating from coconuts with A. guerreronis, but not by odours from coconuts with S. concavuscutum, even when A. guerreronis were present on the same fruit. Fewer N. baraki were recaptured on fruits with both prey or with S. concavuscutum than on fruits with only A. guerreronis. Furthermore, the quality of A. guerreronis from singly and multiply infested coconuts as food for N. baraki did not differ. Concluding, our results suggest that N. baraki does not perform well when S. concavuscutum is present on the coconuts, and the control of A. guerreronis by N. baraki may be negatively affected by the presence of S. concavuscutum.

Estimated crop loss due to coconut mite and financial analysis of controlling the pest using the acaricide abamectin.

Reducing the losses caused by Aceria guerreronis Keifer has been an arduous task for farmers. However, there are no detailed studies on losses that simultaneously analyse correlated parameters, and very few studies that address the economic viability of chemical control, the main strategy for managing this pest. In this study the objectives were (1) to estimate the crop loss due to coconut mite and (2) to perform a financial analysis of acaricide application to control the pest. For this, the following parameters were evaluated: number and weight of fruits, liquid albumen volume, and market destination of plants with and without monthly abamectin spraying (three harvests). The costs involved in the chemical control of A. guerreronis were also quantified. Higher A. guerreronis incidence on plants resulted in a 60 % decrease in the mean number of fruits harvested per bunch and a 28 % decrease in liquid albumen volume. Mean fruit weight remained unaffected. The market destination of the harvested fruit was also affected by higher A. guerreronis incidence. Untreated plants, with higher A. guerreronis infestation intensity, produced a lower proportion of fruit intended for fresh market and higher proportions of non-marketable fruit and fruit intended for industrial processing. Despite the costs involved in controlling A. guerreronis, the difference between the profit from the treated site and the untreated site was 18,123.50 Brazilian Real; this value represents 69.1 % higher profit at the treated site.

Evidence of Amblyseius largoensis and Euseius alatus as biological control agent of Aceria guerreronis.

Amblyseius largoensis (Muma) (Acari: Phytoseiidae) and Euseius alatus De Leon (Acari: Phytoseiidae) are predatory mites that are mostly found on leaves and on the exposed fruit surface of coconut plants. Their morphology hampers the access to the microhabitat occupied by Aceria guerreronis Keifer (Acari: Eriophyidae), the most important pest of coconut fruits throughout the world. However, it was suggested that they can prey on A. guerreronis under natural conditions when this pest leaves its refuge to disperse. Since the trophic interactions between A. largoensis or E. alatus and A. guerreronis are unknown, we compare the frequencies of occurrence of A. largoensis and E. alatus under the bracts of coconut fruits and on coconut leaflets. In addition, because phytoseiids feed by liquid ingestion, we used molecular analysis to confirm the potential role of A. largoensis or E. alatus as predators of A. guerreronis and to assess how fast the A. guerreronis DNA fragment is degradated in the A. largoensis digestive tract. Our study demonstrated that E. alatus was only present on coconut leaflets whereas A. largoensis was found mostly on leaflets and, to a much lesser extent, under the bracts of coconuts. Species-specific ITS primers designed for A. guerreronis were shown to have a high degree of specificity for A. guerreronis DNA and did not produce any PCR product from DNA templates of the other insects and mites associated with the coconut agroecosystem. Based on molecular analysis, we confirmed that the predatory mites, A. largoensis and E. alatus, had preyed on the coconut mite in the field. Overall the predatory mites collected in the field exhibited low levels of predation (26.7% of A. largoensis and 8.9% of E. alatus tested positive for A. guerreronis DNA). The fragment of A. guerreronis DNA remained intact for a very short time (no more than 6 h after feeding) in the digestive tract of A. largoensis.

Endogenous cytokinins in Cocos nucifera L. in vitro cultures obtained from plumular explants.

Auxin induces in vitro somatic embryogenesis in coconut plumular explants through callus formation. Embryogenic calli and non-embryogenic calli can be formed from the initial calli. Analysis of endogenous cytokinins showed the occurrence of cytokinins with aromatic and aliphatic side chains. Fourteen aliphatic cytokinins and four aromatic cytokinins were analysed in the three types of calli and all the cytokinins were found in each type, although some in larger proportions than others. The most abundant cytokinins in each type of callus were isopentenyladenine-9-glucoside, zeatin-9-glucoside, zeatin riboside, isopentenyladenine riboside, dihydrozeatin and dihydrozeatin riboside in decreasing order. Total cytokinin content was compared between the three types of calli, and it was found to be lower in embryogenic calli compared to non-embryogenic calli or initial calli. The same pattern was observed for individual cytokinins. When explants were cultured in media containing exogenously added cytokinins, the formation of embryogenic calli in the explants was reduced. When 8-azaadenine (an anticytokinin) was added the formation of embryogenic calli and somatic embryos was increased. These results suggest that the difference in somatic embryo formation capacity observed between embryogenic calli and non-embryogenic calli is related to their endogenous cytokinin contents.

GA3 stimulates the formation and germination of somatic embryos and the expression of a KNOTTED-like homeobox gene of Cocos nucifera (L.).

The micropropagation of coconut palm has progressed rapidly; yet, there are constraints with regard to the number of somatic embryos formed and their germination. To overcome these, we tested the effect of gibberellic acid and characterized genes of the KNOX family. Gibberellic acid at 0.5 muM increased 1.5-fold the number of calli forming somatic embryos and twofold the number of somatic embryos per callus, calli with germinating embryos and the number of germinating somatic embryos per callus. With regard to the study of KNOX family genes, the complete sequences of two KNOX-like genes were obtained for CnKNOX1 and CnKNOX2. The deduced amino acid sequence of both showed highly conserved domains characteristic of KNOX genes. CnKNOX1 showed high homology with KNOX class I proteins. CnKNOX1 expression was detected throughout the embryogenesis process except in somatic embryos at the pro-globular stage, and was highest in somatic embryos at the coleoptilar stage. No detection of CnKNOX1 expression occurred in calli with aberrant embryos. The addition of gibberellic acid stimulated the expression of CnKNOX1 earlier and the relative expression at all stages was higher. CnKNOX2 expression occurred at all stages peaking at the globular stage, but gibberellic acid treatment decreased the expression. Gene expression was also analyzed in tissues of different organs of adult palms. With CnKNOX1, high level of expression was found in tissues of organs with, but not in those without, meristem, whereas CnKNOX2 expression was detected in tissues with and also in those without meristem.

Detection of a SERK-like gene in coconut and analysis of its expression during the formation of embryogenic callus and somatic embryos.

Somatic embryogenesis involves different molecular events including differential gene expression and various signal transduction pathways. One of the genes identified in early somatic embryogenesis is S OMATIC E MBRYOGENESIS R ECEPTOR-like K INASE (SERK). Cocos nucifera (L.) is one of the most recalcitrant species for in vitro regeneration, achieved so far only through somatic embryogenesis, although just a few embryos could be obtained from a single explant. In order to increase efficiency of this process we need to understand it better. Therefore, the purpose of the present work was to determine if an ortholog of the SERK gene is present in the coconut genome, isolate it and analyze its expression during somatic embryogenesis. The results showed the occurrence of a SERK ortholog referred to as CnSERK. Predicted sequence analysis showed that CnSERK encodes a SERK protein with the domains reported in the SERK proteins in other species. These domains consist of a signal peptide, a leucine zipper domain, five LRR, the Serine-Proline-Proline domain, which is a distinctive domain of the SERK proteins, a single transmembrane domain, the kinase domain with 11 subdomains and the C terminal region. Analysis of its expression showed that it could be detected in embryogenic tissues before embryo development could be observed. In contrast it was not detected or at lower levels in non-embryogenic tissues, thus suggesting that CnSERK expression is associated with induction of somatic embryogenesis and that it could be a potential marker of cells competent to form somatic embryos in coconut tissues cultured in vitro.