Establishment Of Tissue Culture And Hairy Root Production Systems For Solenostemon Scutellarioides (L.) Codd

In vim propagation of S. scutellarioEdes was canied out by culturing shoot tip explants onto LS, MS and 65 basal medium supplemented with different concentrations of BAP. Comparative analysis of the different types of basal media showed that the LS basal medium supplemented with 8.88 pM BAP was suitable for multiple shoots formation on S. scutellarioides (- 13 shoots/explant). Addition of higher concentrations of BAP (35.5 pM) inhibited multiple shoots and induced abnormal plantlet formation in this species. Culturing shoot tip explants on 65 medium supplemented with 4.52 pM BAP and 2.26 pM 2,4-D produced pink friable callus. On the other hand, culturing leaf explants on MS and 65 media both supplemented with 4.52 pM 2,4-D and 0.46 pM 2,4-D, produced yellowish and grayish friable calli, respectively. Following calli induction, cell suspension culture of S. scu2eIlarirw'des was successfully initiated on MS medium supplemented with 2.26 pM 2,4-D and 0.47 pM kinetin. The cell suspension required only half the amount of 2,443 iii required for calli induction on solid medium (MS + 4.52 pM 2,4-D + 0.46 pM kinetin). In an effort towards achieving genetically stable plant tissue culture material, for the future indudion of valuable secondary metabolite, adventitious roots and hairy roots were induced from S. scufellarioides explants. Quantitative and qualitative assessments of the biomass producing from each of the culture method were analyzed. Adventitious root cultutes in S. sartellarioides leaves were induced by placing explants onto MS basal medium supplemented with different concentrations of auxins (NAA, IBA, IAA) and cytokinin (kinetin). Comparative analysis of the mots enhanced showed that MS medium supplemented with 5.0 pM IBA produced rapidly growing adventitious roots. The presence of cytokinin (kinetin) was inhibiting to adventitious root formation in S. SCUfellarioides. Supplementing MS medium with auxin either IBA or NAA was enough to induce adventitious root formation. Hairy roots of S. scutellarioides were induced by inoculation of leaf explants with A. fiizogenes strains TR 105, LBA 9402,8196 and ATCC 15834. These strains showed different abilities to induce hairy root formation in the leaf explants. Assessment of the plant susceptibility to the different A. rhizogenes strains showed that the strains ATCC 15834, TR 105, LBA 9402, and 81 96 produced 56.3%, 25.5%, 21.5%, and 13.8% transformation efficiencies, respectively. Acetosyringone was found to be useful for enhancement of hairy root production in S. s c u t d I 8

Molecular Characterization and Functional Analysis of Selected Expressed Sequence Tags from Oil Palm Cell Suspension Culture

A large quantity of Expressed sequence tags (ESTs) are available from various cDNA libraries of oil palm. The information from oil palm EST databases has been utilized to identify several interesting transcripts that are upregulated in cell suspension culture for molecular characterization and functional analysis. The first part of this study was to carry out the molecular characterization of selected ESTs of oil palm cell suspension culture which were Eg583 (Accession number: EU795363), Eg707 (Accession number: FJ196136) and EgHAD (Accession number: FJ196137). The Eg583 sequence is highly similar to an unknown protein from rice. This predicted protein might be a transcription factor due to the presence of SIN3 domain and motifs of casein kinase II phosphorylation. The expression of this gene was not detectable in all tested tissues. This gene might be a member of a multigene family in the oil palm genome. The Eg707 sequence is highly similar to an unknown protein from Arabidopsis and might be a putative nuclear protein. Its amino acid sequence contains a Ald-Xan-dh-C2 domain that may be involved in ABA biosynthesis. Eg707 might be present as a single copy gene in the oil palm genome and its transcripts were highly expressed in tissue cultured materials compared to vegetative tissues. Eg707 might have a role during oil palm somatic embryogenesis or at very early stage of embryo development. The EgHAD sequence is similar to a putative haloacid dehalogenase (HAD) superfamily hydrolase from monocots and phosphate hydrolase from dicots. However, the phylogenetic relationship of EgHAD is closer to monocots than dicots. EgHAD might be a member of a multigene family gene in the oil palm genome. It was highly expressed in leaves and meristem but lower expression was found in roots, female flowers, non-embryogenic and embryogenic calli in comparison to the oil palm cell suspension culture. Functional analysis was carried out in rice by over-expressing Eg707 and EgHAD, driven by a constitutive double Cauliflower Mosaic Virus 35S promoter. The constructs were made using the gateway technology with clonase (Invitrogen, USA). Transgenic plants over-expressing Eg707 protein had small sized, rolled and erect leaves, less tillers, empty seeds and higher total chlorophyll content. The phonotypes of these and the presence of Xan-dh-C2 domain in Eg707 protein, strongly suggest its involvement in ABA biosynthesis, particularly during somatic embryogenesis. Functional analysis of Eg707 through RNAi-mediated gene silencing was unsuccessful since the T1 seeds failed to germinate. Over-expression of EgHAD gene in rice produced more lateral roots and tillers than the wild type plants. However, it also reduced plant size, produced empty seeds and many tiny seeds which were not found in wild type plants. The suppression of the EgHAD orthologues in rice did not show any changes in the phenotype. EgHAD might be a metabolic protein involved in phosphate starvation mechanism and its expression might be necessary for seed germination

Overexpression of the oil palm (Elaeis guineensis Jacq.) TAPETUM DEVELOPMENT1-like Eg707 in rice affects cell division and differentiation and reduces fertility

The functional analysis of the TAPETUM DEVELOPMENT1-like analog Eg707 of oil palm was carried out in rice by over-expressing Eg707 under the control of a double cauliflower mosaic virus 35S promoter. Ectopic expression of Eg707 in rice induced dark green and matured compact brownish calli compared to pale wild type and negative control calli. Regenerated transgenic rice plants exhibited a reduction in organ size and plant height, rolled, erect leaves, less tillers, increased chlorophyll content, and reduced fertility with smaller green seeds. At the molecular level Eg707 overexpression caused an increase in the transcription of SAPK9, a SnRK2 protein kinase family member that is activated by ABA and hyperosmotic stress. Together, the results show that ectopic Eg707 expression influences cell division and differentiation, presumably via altered hormone homeostasis

Profiling the differentially expressed genes in two rice varieties during rapid grain-filling stages

Grain filling is an important agronomic trait, which directly affects the final yield of rice. Partially filled and empty rice grains are among the factors that limit the yield of MR219, one of the highest yielding rice varieties in Malaysia. In this study, the NSF 20 K rice oligonucleotide array, which contains 20,000 70-mer oligonucleotide probes, was used for direct comparison of the transcriptomes of MR219 and MR84 (a rice variety that has higher percentage of filled grains compared to MR219), during rapid grain-filling period at 5 and 10 days after fertilization (DAF). A total of 155 and 233 genes were differentially expressed in MR219 compared to MR84 at 5 and 10 DAF, respectively; and 9 of these expression ratios were tested using quantitative real-time RT PCR. Among the differentially expressed genes identified were those encoding hexokinase, various sugar transporters, GSDL-like lipase/acylhydrolase, brassinosteroid-insensitive 1-associated receptor kinase 1 precursor and homeobox protein GLABRA2, which were analyzed by real-time RT PCR in this study. The differences demonstrated by these genes in their transcript levels and profiles, between the two rice varieties understudied at different stages of grain filling may contribute to the formulation of hypotheses toward the understanding of poor percentage of filled grains in MR219

A novel transcript of oil palm (Elaeis guineensis Jacq.), Eg707, is specifically upregulated in tissues related to totipotency

In this study, we report the molecular characterization of clone Eg707 isolated from cell suspension culture of the oil palm. The deduced polypeptide of clone Eg707 is highly similar to an unknown protein from Arabidopsis thaliana. The presence of an Ald-Xan-dh-C2 superfamily domain in the deduced protein sequence suggested that Eg707 protein might be involved in abscisic acid biosynthesis. Eg707 might be present as a single copy gene in the oil palm genome. This gene is highly expressed in tissue cultured materials compared to vegetative and reproductive tissues, suggesting a role of this gene during oil palm somatic embryogenesis or at the early stages of embryo development. Expression analysis of Eg707 by RNA in situ hybridization showed that Eg707 transcripts were present throughout somatic embryo development starting from proembryo formation at the embryogenic callus stages till the maturing embryo stages. Since proembryo formation within the embryogenic callus is one of the first key factors in oil palm somatic embryo development, it is suggested that Eg707 could be used as a reliable molecular marker for detecting early stage of oil palm somatic embryogenesis

Safety and efficacy of fluoxetine on functional outcome after acute stroke (AFFINITY): a randomised, double-blind, placebo-controlled trial

Background Trials of fluoxetine for recovery after stroke report conflicting results. The Assessment oF FluoxetINe In sTroke recoverY (AFFINITY) trial aimed to show if daily oral fluoxetine for 6 months after stroke improves functional outcome in an ethnically diverse population. Methods AFFINITY was a randomised, parallel-group, double-blind, placebo-controlled trial done in 43 hospital stroke units in Australia (n=29), New Zealand (four), and Vietnam (ten). Eligible patients were adults (aged ≥18 years) with a clinical diagnosis of acute stroke in the previous 2–15 days, brain imaging consistent with ischaemic or haemorrhagic stroke, and a persisting neurological deficit that produced a modified Rankin Scale (mRS) score of 1 or more. Patients were randomly assigned 1:1 via a web-based system using a minimisation algorithm to once daily, oral fluoxetine 20 mg capsules or matching placebo for 6 months. Patients, carers, investigators, and outcome assessors were masked to the treatment allocation. The primary outcome was functional status, measured by the mRS, at 6 months. The primary analysis was an ordinal logistic regression of the mRS at 6 months, adjusted for minimisation variables. Primary and safety analyses were done according to the patient's treatment allocation. The trial is registered with the Australian New Zealand Clinical Trials Registry, ACTRN12611000774921. Findings Between Jan 11, 2013, and June 30, 2019, 1280 patients were recruited in Australia (n=532), New Zealand (n=42), and Vietnam (n=706), of whom 642 were randomly assigned to fluoxetine and 638 were randomly assigned to placebo. Mean duration of trial treatment was 167 days (SD 48·1). At 6 months, mRS data were available in 624 (97%) patients in the fluoxetine group and 632 (99%) in the placebo group. The distribution of mRS categories was similar in the fluoxetine and placebo groups (adjusted common odds ratio 0·94, 95% CI 0·76–1·15; p=0·53). Compared with patients in the placebo group, patients in the fluoxetine group had more falls (20 [3%] vs seven [1%]; p=0·018), bone fractures (19 [3%] vs six [1%]; p=0·014), and epileptic seizures (ten [2%] vs two [<1%]; p=0·038) at 6 months. Interpretation Oral fluoxetine 20 mg daily for 6 months after acute stroke did not improve functional outcome and increased the risk of falls, bone fractures, and epileptic seizures. These results do not support the use of fluoxetine to improve functional outcome after stroke

Improvement of Glutinous Corn and Watermelon Yield by Lime and Microbial Organic Fertilizers

Background. The characteristics of acid soil, often low pH and high toxicity, affect the growth and yield of plants. Aims. This study evaluates the effects of supplemented lime and microbial organic fertilizer on glutinous corn (Zea mays) and watermelon (Citrullus lanatus) yield, yield components, and economic efficiency on acid-sulfate soils. Materials and Methods. Two experiments were carried out in Phung Hiep District, Hau Giang Province, as a typical acid-sulfate soil area. The randomized complete block with four treatments and three replicates was designed for the experiment, in which supplemented fertilizers were 800 kg of lime/ha; 2,000 kg of microbial organic fertilizer/ha; and 800 kg of lime in combination with 2,000 kg of microbial organic fertilizer/ha, and treatment as farmer dose (FFT), without lime and microbial organic fertilizer. Results. As a result, using lime combined with microbial organic fertilizer increased the yield compared to using only lime or microbial organic fertilizer. Besides, the incomes increased to 12.0% and 13.8%, respectively, compared to farmer recommendations. Conclusion. To improve the yield of glutinous corn or watermelon and income on acid-sulfate soils, lime should be applied at 800 kg combined with 2,000 kg of microbial organic fertilizer/ha

Assessment of hairy roots induction in Solenostemon scutellarioides leaves by different strains of Agrobacterium rhizogenes

Hairy roots of Solenostemon scutellarioides were induced by inoculation of leaf explants with Agrobacterium rhizogenes strains TR 105, LBA 9402, 8196 and ATCC 15834. These strains showed different abilities to induce hairy root formation on the leaf explants. Assessment of the plant’s susceptibility to the different strains of A. rhizogenes showed that strains ATCC 15834, TR 105, LBA 9402, and 8196 produced 56.3, 25.5, 21.5 and 13.8% transformation efficiencies, respectively. Acetosyringone was found to be useful for the enhancement of hairy roots formation in S. scutellarioides

Impact of saltwater intrusion on relation to fruit growing households in Ben Tre, Vietnam

Purpose. Saltwater intrusion affected seriously the livelihood of mangosteen growers in Ben Tre, a coastal province in the Vietnamese Mekong Delta in 2016. This study assesses how saltwater intrusion influenced the livelihood vulnerability of the mangosteen households and communities. Methodology / approach. This study used the United Nations’ Livelihood Vulnerability Index (LVI) and a similar index from the Intergovernmental Panel on Climate Change (LVI-IPCC) using data from 196 interviewed mangosteen growers in Cho Lach district, Ben Tre province in 2016 and 2018. A total of twenty-five (25) indicators are applied to calculate the two indices, using data as exposure to saltwater intrusion and natural hazards (5), socio-demographic factors (11), and sensitivity of health, finance, and source of water for domestic use (9). Results. The analysis results show that mangosteen-growing households had a medium vulnerability in 2016 and 2018 based on both indices. However, they were extremely vulnerable due to saltwater intrusion in 2016, and a high household percentage had a moderate value in the vulnerability index. They did not experience how to respond to saltwater intrusion and received a late warning on saltwater intrusion from the local authorities and media. However, mangosteen-growing households had a quick adaptive behavior to shift to new crops to improve their income in 2018. Originality / scientific novelty. The study provides a set of indicators to assess the vulnerability of this low-vulnerable area based on past studies as well as the actual situation of the study area in 2016 and 2018 to determine what factors influenced the main components, LVI and LVI – IPCC. These indicators represented the characteristics of the rural households in Vietnam and other Asian countries. This research also indicated how LVI and LVI – IPCC results differently and when they should be used. The analysis identified the factors influencing the LVI and LVI – IPCC by levels of saltwater intrusion in different years of 2016 and 2018, two different severe and less severe sites, adaptive and non-adaptive households, and their interaction. Practical value / implications. LVI and LVI-IPCC are good indicators for local and regional assessments on how saltwater intrusion and natural hazards affect households’ and communities’ livelihood and thereby how to guide them to respond promptly in such cases

Molecular characterization of an unknown protein (Acc. No. EU795363) from the ESTs of oil palm (Elaeis guineensis Jacq.) cell suspension culture.

A large quantity of ESTs is available from various cDNA libraries of oil palm. The information from oil palm EST databases has been utilized to identify several interesting sequences for molecular characterization. In this study, we report molecular characterization of clone 583 (Acc. No. EU795363) isolated from cell suspension culture of oil palm. This clone is predicted to encode a single major open reading frame for a polypeptide of 177 amino acids with a predicted molecular mass of 19.6 kDa. The predicted amino acid sequence does not contain any signal peptide and transmembrane region. Based on Kyte-Doolittle hydropathy profile, this protein is predicted to be a soluble protein. The predicted ORF of clone 583 was 59% identical to an amino acid sequence of an unknown protein from Oryza sativa (Acc. BAD25663). Southern analysis showed that this clone might be a member of a multigene family in the oil palm genome. Gene expression study by real time quantitative RT-PCR showed that transcripts of clone 583 might be present in low abundance