Resumo -O objetivo deste trabalho foi avaliar genótipos de capim-elefante (Pennisetum purpureum Schum.) quanto à resistência à cigarrinha-das-pastagens (Mahanarva spectabilis). Para avaliação da antibiose, aos trinta dias após o plantio, cada planta foi infestada com seis ovos próximos à eclosão, em delineamento inteiramente casualizado, com 30 genótipos e dez repetições. Quarenta e cinco dias após a eclosão das ninfas, avaliou-se a porcentagem de sobrevivência do inseto-praga nos diferentes genótipos. Para avaliação da não-preferência, foram quantificados, quinzenalmente, o número e tamanho de ninfas por vaso, em plantas mantidas em casa de vegetação, onde adultos de M. spectabilis eram periodicamente liberados, em delineamento inteiramente casualizado, com três repetições por genótipo, em dois períodos de amostragem. Os genótipos Cameroon de Piracicaba, Pioneiro, Cuba 169, Santa Rita, Mineiro Ipeaco, Mercker Comum de Pinda e CNPGL 96-27-3 foram selecionados quanto à resistência, pelo mecanismo de antibiose. O número e o tamanho médio das ninfas variaram significativamente em razão do genótipo de capim-elefante, no estudo da não-preferência. Os genótipos Roxo de Botucatu e Pioneiro são candidatos à testemunha suscetível e resistente, respectivamente, pelo mecanismo de antibiose, e os genótipos Cameroon e Cameroon Piracicaba são promissores pelo mecanismo de não-preferência.Termos para indexação: Pennisetum purpureum, Mahanarva spectabilis, resistência a pragas, forrageiras, antibiose, não-preferência. Selection of elephant grass genotypes for resistance to spittlebugAbstract -The objective of this work was to evaluate genotypes of elephant grass (Pennisetum purpureum Schum.) regarding resistance to the spittlebug Mahanarva spectabilis. For antibiosis evaluation, each plant was infested with six eggs near hatching, thirty days after planting, in a completely randomized design with 30 genotypes and ten repetitions. Forty-five days after the nymphs hatched, the insect survival rate was evaluated on the different genotypes. For nonpreference mechanism evaluation, the size and number of the nymphs per pot were assessed every 15 days, in plants kept at greenhouse, where adults of M. spectabilis were periodically released, in a completely randomized design, with three repetitions per genotype in two sampling periods. Cameroon de Piracicaba, Pioneiro, Cuba 169, Santa Rita, Mineiro Ipeaco, Mercker Comum de Pinda and CNPGL 96-27-3 genotypes were selected for resistance, by antibiosis mechanism. In the study of the nonpreference mechanism, the number and average size of the nymphs varied significantly as a function of the elephant grass genotype. Roxo de Botucatu and Pioneiro genotypes are, respectively, susceptible and resistant check candidates by the antibiosis mechanism, and Cameroon and Cameroon Piracicaba genotypes are promising by the nonpreference mechanism.
The present study evaluated spermatozoal capacitation in Farfantepenaeus paulensis. This process has direct applications in aquaculture, and it consists of the ionic, biochemical and morphological changes during the period that the spermatophore is stored or adhered to the thelycum. These changes make the spermatozoa capable of fertilization. The morphological changes of spermatozoal capacitation have been previously investigated only in sicyoniids and open-thelycum penaeids. Thus, this study is the first morphological account of a closedthelycum penaeid. F. paulensis broodstock were captured offshore in southern Brazil, and spermatophores were collected from the terminal ampoule and from the thelycum of sexually mature females that had naturally copulated. The ultrastructure of the spermatozoal capacitation was investigated via transmission electron microscopy. Spermatozoa of F. paulensis show the following changes related to capacitation: (1) the chromatin became less condensed; (2) the acrosomal region became more electron-dense (3) the acrosomal cap became less concave; and (4) the subacrosomal region became much more electron-dense. These results demonstrate that the morphological changes in capacitated spermatozoa of F. paulensis are similar to those previously reported for open-thelycum penaeids, i.e., Litopenaeus species. Further studies on the capacitation process are required, especially to evaluate the interaction between biochemical and morphological changes. Such research could be useful for developing biotechnologies that will allow spermatozoal induction without storing the spermatophore in the thelycum and therefore allow in vitro larval production.
The method usually employed to stimulate gonadal maturation and spawning of captive shrimp involves unilateral eyestalk ablation, which results in the removal of the endocrine complex responsible for gonad-inhibiting hormone (GIH) synthesis and release. In the present study, RNAi technology was used to inhibit transcripts of GIH in Litopenaeus vannamei females. The effect of gene silencing on gonad development was assessed by analyzing the expression of GIH and vitellogenin, respectively, in the eyestalk and ovaries of L. vannamei females, following ablation or injection with dsRNA-GIH, dsRNA-IGSF4D (non-related dsRNA), or saline solution. Histological analyses were performed to determine the stage of gonadal development and to assess the diameter of oocytes throughout the experimental procedure. Only oocytes at pre-vitellogenesis and primary vitellogenesis stages were identified in females injected with dsRNA-GIH, dsRNA-IGSF4D, or saline solution. Oocytes at all developmental stages were observed in eyestalk-ablated females, with predominance of later stages, such as secondary vitellogenesis and mature oocytes. Despite achieving 64, 73, and 71% knockdown of eyestalk GIH mRNA levels by 15, 30, and 37 days post-injection (dpi), respectively, in dsRNA-GIH-injected females, the expected increase in ovary vitellogenin mRNA expression was only observed on the 37th dpi. This is the first report of the use of RNAi technology to develop an alternative method to eyestalk ablation in captive L. vannamei shrimps.
Shrimp farming worldwide is based on a similar technological package, characterized by three phases: Controlled Reproduction, Larvae Culture, and Grow-out Culture. This basic aquaculture package uses broodstock animals, with different levels of fundamental genetic selection, induced to mature and reproduce based on unilateral eyestalk ablation. This review identified ten reproduction-related research subjects that can improve the shrimp industry based on basic scientific knowledge and four levels of application: eyestalk ablation alternatives, larvae production, product protection, and grow-out yield improvement. Species-specificity must be considered in developing biotechnology solutions. Alternatives to eyestalk ablation for controlling ovarian maturation are based on neurotransmitter regulation in Litopenaeus; however, the environmental impact of this approach has to be evaluated. Maturation by RNAi requires further evaluation, and maturation pheromones have not been explored. Sex reversal, hybridization, in vitro fertilization, and seedstock cryopreservation of penaeids require fundamental research. Triploid culture and genetic selection can be applied to some species for product protection; however, for L. vannamei, triploidization is not practical yet.
Braga, A.L., Nakayama, C.L., Suita de Castro, L.A. and Wasielesky, W. 2011. Spermatozoa ultrastructure of the pink shrimp Farfantepenaeus paulensis (Decapoda: Dendrobranchiata). —Acta Zoologica (Stockholm) 00: 1–6. The spermatozoa ultrastructure of the pink shrimp Farfantepenaeus paulensis was investigated in this morphological study. Spermatophores and spermatozoa were analyzed by electron microscopy. The pink shrimp spermatophore is divided into two regions: the appendage and the spermatophore main body, where spermatozoa are grouped in a spermatic mass. Pink shrimp spermatozoa are unistellate and are composed of main body and single spike. The spermatozoa body comprises a perinuclear cytoplasmic band, nucleus, acrosomal cap, and subacrosomal region. The spermatozoa cell mean total length was 10.71 μm, the mean body diameter was 5.56 μm, and the mean spike length and diameter were 5.15 μm and 0.85 μm, respectively.
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