Effects of different quality of soil mixture on growth development of an important medicinal plant, Boesenbergia rotunda

Growth and morphological development of Boesenbergia rotunda grown in different soil mixture were considered to determine the suitable growing media for the species. B. rotunda or fingerroot ginger is a highly important medicinal plant belonging to Zingiberaceae family. The rhizome and fingerroot structure of this species contains several bioactive compounds with various functional pharmaceutical activities. It can be vegetatively propagated through cutting rhizome and shows slow growth rate. This study provided some analysis and informative data on how the three types of typical soil (red soil (RS), black soil (BS) and sand (SS) can give important influences on the morphological and physiological development of the species. Fourteen different types of soil mixture with different mix ratio and quality were used as a growth medium. The effects of these treatments were implied based on the growth rate, evaluation in biomass quality of shoots, rhizomes and fingerroots, and photosynthetic pigment analysis. The highest quality growth of B. rotunda was established in the medium containing high percentage of RS and BS with low of SS. The growth rate of plant and photosynthetic pigment concentration were increased in the medium containing a high percentage of RS (50-100%) and BS (50-100%). The presence of a high percentage of BS in the medium was also significantly increased the biomass production of rhizome and fingerroots. The soil mixture might not make adverse effects on the shoots biomass except in medium containing more than 50% of SS. The physical characteristics (bulk density, porosity, water holding capacity and electrical conductivity) of the soil mixture were studied to determine the quality of an optimum combination of the growing medium. The synthetically evaluation index of the plant showed that the different type of soil mixture has a significant effect on growth development of B. rotunda that necessary in industrialization cultivation study

Thymoquinone enhances sperm DNA integrity in nicotineinduced infertile male rats

Purpose: To assess the effects of thymoquinone (TQ) on the integrity of sperm DNA in nicotineinduced sperm impairment in rats. Methods: Adult male Sprague Dawley rats were randomized into four equal groups: control group received normal saline orally for 60 days; nicotine group was subcutaneously injected with 5 mg/kg/day nicotine for 30 days and then given normal saline for the next 30 days; TQ group was given normal saline for 30 days followed by TQ at 5 mg/kg/day for 30 days; and nicotine-TQ group received 5 mg/kg of nicotine for 30 days and 5mg/kg of TQ for another 30 days. Sperm DNA breakages were evaluated using Comet assay. The expression levels of protamine 1 (PT1) and transition nuclear protein 2 (Tnp2) genes which are essential for the proper compaction of the sperm DNA were analyzed by quantitative polymerase chain reaction (qPCR). Results: Thymoquinone significantly decreased DNA fragmentation in the sperm of nicotine-treated rats. However, there was no change in PT1 gene expression. Tnp2 was downregulated in the nicotine group and slightly upregulated in nicotine-TQ group (p < 0.05). Conclusion: The results demonstrate the potential benefits of TQ in improving sperm DNA quality of nicotine-induced male infertility

RT-qPCR profiling of pathogenesis related genes in Musa acuminata cv. 'Berangan' seedlings challenged with Fusarium oxysporum F. SP. Cubense tropical race 4

The expression profile of pathogenesis related genes are signatures of an infection response in plant cells. Pathogenic infections can increase or reduce gene expression in a plant system in a relatively specific pattern. These expression patterns can be used as standards in pathogenicity studies and, where phenotypic expression is normally used to gauge a plants response to infection, it could additionally present a more rapid and early screening reference tool. Three genes: catalase (CAT), pathogen related protein (PR10), and phenylalanine ammonia (PAL) all implicitly implicated in the plant disease response pathway were targeted for analysis during the infection of Fusarium oxysporum f. sp. cubense tropical race 4 (FOCR4) in banana Musa acuminata cv. Berangan seedlings after a standard challenge under growth room conditions. Distinct patterns of gene expression were observed at three infection time points by real time expression analysis. There was a sequential 10-fold reduction in expression for the PR gene while, the PAL and CAT genes were both upregulated. These results present a set of reference genes that could be used for screening of a plant’s response to Fusarium before the onset of symptoms

Elucidation of Musa acuminata cv. Berangan root infection by FOC (Tropical Race 4) by RNA sequencing and analysis

Musa acuminata cv. Berangan (AAA) is a type of banana locally grown in Malaysia. These bananas as well as Musa acuminata cv. Cavendish (AAA) are also facing a major threat from a typical soil borne fungus identified as Fusarium oxysporum f. sp. cubense race 4 (FocR4). Its characteristics as a complex pathogen manifesting as subtypes or races are the main reasons its infections are difficult to control. Genome sequence availability of the double haploid Musa acuminata originating from Pahang has become very useful to analyse RNA-seq reads and to identify the transcriptome profile of the host response between different groups was accomplished using RNA-Seq technology based on the Illumina HiSeqTm 2000 platform. Three sets of libraries derived from infected and mock infected plants (experimental groups) between different time points (0, 48, 96 h) shows over forty million reads were generated, each corresponding to coverage of >4,000,000,000 to <8,000,000,000 bases. About 0.10-66% reads were mapped to Musa acuminata DH Pahang genome sequence. This study provides the statistical analysis of the sequence reads. Based on this information, further analysis on gene expression patterns influenced by Foc race infection within the tested groups and time points will help in the understanding of the host pathogenic responses. In future discovery on many new genes for diagnosis of plant infection could be achieved through excessive transcriptomic data

The effect of pathogenesis-related 10 (Pr-10) gene on the progression of fusarium wilt in Musa acuminata cv. Berangan

PR-10 is a member of pathogenesis-related (PR) genes elicited by the plant’s defense mechanism during pathogen attack. Elevated expression of PR-10 upon different pathogen invasions has been observed in many plant species suggesting its role as an anti-bacterial, anti-viral and anti-fungal gene. However, the effect of PR-10 in mitigating the infection of Fusarium oxysporum f. sp. cubense (Foc), the causal agent of Fusarium wilt in banana has not been reported. In this study, the coding sequences of PR-10 gene isolated from Foc resistant Musa acuminata ssp. malaccensis (MaPR-10) were integrated into a local Foc susceptible commercial banana cultivar, Berangan via co-cultivation of embryogenic cell suspension and Agrobacterium tumefaciens. Out of 17 putative transgenic lines established, 11 of them positively harbored MaPR-10. Among these, Line-19 plantlets showed the most rapid in-vitro propagation and successfully over-expressed the transgene. Following a nursery challenge experiment with a virulent Foc race 4 (CI HIR) isolate, about 30% of Line-19 plants showed a one-week delay in disease progression when compared to the untransformed controls. From the final evaluation performed in the 5th week-post-inoculation, the leaf symptoms index (LSI) and rhizome discoloration index (RDI) of Line-19 was 3.4 and 6.1, respectively, indicating the disease had progressed. The findings of this study enrich the current existing knowledge on the roles of PR-10 in combating fungal disease in plants

Standardized bioassays: an improved method for studying Fusarium oxysporum f. sp. cubense race 4 (FocR4) pathogen stress response in Musa acuminata cv. ‘Berangan’

To date, there is no standardized Fusarium bioassay protocol established owing partly to the wide variety of Fusarium oxysporum f. sp. cubense (Foc) isolates and banana cultivars present. Thus, validation of the infection parameters is deemed essential prior to each bioassay experiment. In the current study, a simple standardized workflow was developed based on available assays for testing Fusarium wilt disease response in Musa acuminata using M. acuminata cv. ‘Berangan’ of tissue-culture origin as a model. The phenotypic assays were able to detect external disease symptoms less than one week post-inoculation, while the molecular approach using RT-qPCR identified differential expression of catalase (CAT), pathogenesis-related 10 (PR10), phenylalanine ammonia-lyase (PAL) and xylanase (XYL) genes as early as day 0. The transcript levels of PR10 and XYL fluctuated over 4 days of Foc Race 4 (FocR4 C1 HIR isolate) infection while the expression of CAT steadily increased over time. In contrast, PAL was highly upregulated at 2 days post-inoculation. These signature changes suggest that all genes tested might be involved in the early defense response of ‘Berangan’ plants against FocR4 infection. ‘Berangan’ cultivar was found to be highly susceptible to Foc Race 4 (C1 HIR isolate) with leaf symptoms index (LSI) and rhizome discoloration index (RDI) scores of 4.257 and 5.971, respectively. The procedure elaborated in this study can be used as a reference Foc bioassay for reproducible and comparable results possibly across cultivars and test isolates due to its simple steps aided by integration of phenotypic and molecular approach

Immunocytochemical evidence of histamine 1 and histamine 2 receptors on mice sperm

632-639Histamine is a biogenic amine which is synthesised by L-histidine decarboxylase enzyme (HDC). The histamine 1 and 2 antagonist administrations have been highly reported to cause detrimental effect on sperm parameters, which arisen the speculation of histamine 1 (H1R) and histamine 2 (H2R) receptors might be present in sperm. The present study was aimed to provide evidence on the localisation of H1R and H2R on mice sperm through immunocytochemistry. The sperm was harvested from cauda epididymis. After one hour of incubation, sperm suspension was smeared onto a poly-lysine-coated slide and allowed to dry before fixation and permeabilisation processes. The primary antibody encoded for receptors was exposed to the fluorescently tagged antibody; fluorescein isothiocyanate (FITC) conjugate followed by nuclear staining with 4, 6-diamino-2-phenylindole dihydrochloride (DAPI). The testis, stomach, and skin were used as the positive controls. Our data showed that both receptors have been expressed on the midpiece and acrosome of mice. The present result was the first discovery of the presence and immunolocalisation of H1R and H2R on mice sperm. Therefore, present study proposes that these receptors could be involved in calcium regulatory mechanism and protein phosphorylation which are responsible for fertilisation-related processes

Immunocytochemical evidence of histamine 1 and histamine 2 receptors on mice sperm

Histamine is a biogenic amine which is synthesised by L-histidine decarboxylase enzyme (HDC). The histamine 1 and 2 antagonist administrations have been highly reported to cause detrimental effect on sperm parameters, which arisen the speculation of histamine 1 (H1R) and histamine 2 (H2R) receptors might be present in sperm. The present study was aimed to provide evidence on the localisation of H1R and H2R on mice sperm through immunocytochemistry. The sperm was harvested from cauda epididymis. After one hour of incubation, sperm suspension was smeared onto a poly-lysine-coated slide and allowed to dry before fixation and permeabilisation processes. The primary antibody encoded for receptors was exposed to the fluorescently tagged antibody; fluorescein isothiocyanate (FITC) conjugate followed by nuclear staining with 4, 6-diamino-2-phenylindole dihydrochloride (DAPI). The testis, stomach, and skin were used as the positive controls. Our data showed that both receptors have been expressed on the midpiece and acrosome of mice. The present result was the first discovery of the presence and immunolocalisation of H1R and H2R on mice sperm. Therefore, present study proposes that these receptors could be involved in calcium regulatory mechanism and protein phosphorylation which are responsible for fertilisation-related processes