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Phalaenopsis amabilis (L.) Blume orchid is an Indonesian national flower. The number of these orchids in their natural habitat is very limited, therefore plant propagation efforts are needed. One of the promising methods is plant propagation by inserting embryo gene AtRKD4 from a model plant Arabidopsis thaliana into the orchid genome to produce many somatic embryos. From previous research, we have obtained 28 plant P. amabilis transformants carrying the AtRKD4 gene, however, it was unknown whether these plants have normal phenotypes and growth similar to their parents. Therefore, descriptions on growth and morphology are needed. This research aimed to evaluate the phenotype of P. amabilis carrying 35S::AtRKD4 the transformants grown in greenhouse. To achieve it, AtRKD4 gene integration stability on transformants genome was analyzed. Morphology and cross-section anatomy structure on transformant and non-transformant plantswere described. The stability of AtRKD4 gene integration in the plant genome was confirmed by amplification of the AtRKD4 gene from genomic DNA with Polymerase Chain Reaction (PCR) using a specific primer for AtRKD4 and ACTIN genes as the internal control. The quantitative data from morphology and anatomy measurements were analyzed statistically using ANOVA. The results showed that AtRKD4 was stably integrated into the genome of P. amabilis transformants and all transformant plants showed similar morphology and anatomy characteristics as non-transformant plants. The AtRKD4 embryo gene was stably integrated into the orchid genome and the transformant plants grow normally without significant changes in phenotype.

Background Plants have the unique capability to form embryos from both gametes and somatic cells, with the latter process known as somatic embryogenesis. Somatic embryogenesis (SE) can be induced by exposing plant tissues to exogenous growth regulators or by the ectopic activation of embryogenic transcription factors. Recent studies have revealed that a discrete group of RWP-RK DOMAIN-CONTAINING PROTEIN (RKD) transcription factors act as key regulators of germ cell differentiation and embryo development in land plants. The ectopic overexpression of reproductive RKDs is associated with increased cellular proliferation and the formation of somatic embryo-like structures that bypass the need for exogenous growth regulators. However, the precise molecular mechanisms implicated in the induction of somatic embryogenesis by RKD transcription factors remains unknown. Results In silico analyses have identified a rice RWP-RK transcription factor, named Oryza sativa RKD3 (OsRKD3), which is closely related to Arabidopsis thaliana RKD4 (AtRKD4) and Marchantia polymorpha RKD (MpRKD) proteins. Our study demonstrates that the ectopic overexpression of OsRKD3, which is expressed preferentially in reproductive tissues, can trigger the formation of somatic embryos in an Indonesian black rice landrace (Cempo Ireng) that is normally resistant to somatic embryogenesis. By analyzing the transcriptome of induced tissue, we identified 5,991 genes that exhibit differential expression in response to OsRKD3 induction. Among these genes, 50% were up-regulated while the other half were down-regulated. Notably, approximately 37.5% of the up-regulated genes contained a sequence motif in their promoter region, which was also observed in RKD targets from Arabidopsis. Furthermore, OsRKD3 was shown to mediate the transcriptional activation of a discrete gene network, which includes several transcription factors such as APETALA 2-like (AP2-like)/ETHYLENE RESPONSE FACTOR (ERF), MYB and CONSTANS-like (COL), and chromatin remodeling factors associated with hormone signal transduction, stress responses and post-embryonic pathways. Conclusions Our data show that OsRKD3 modulates an extensive gene network and its activation is associated with the initiation of a somatic embryonic program that facilitates genetic transformation in black rice. These findings hold substantial promise for improving crop productivity and advancing agricultural practices in black rice.

The number of incidences and deaths from cancer is increasing every year around the world. However, currently available cancer treatment methods still have weaknesses, the discovery of cancer drugs is still very important. The purpose of this study was to determine the effect of the level of public knowledge about breast cancer with BSE behavior on housewives in Negeri Lima Village, Maluku Province. The design of this research is descriptive analytic with a cross-sectional approach. The study was conducted in May 2022. The population in this study were residents of the village of Negeri Lima. Sampling using the method with 50 respondents. The tool used in this research is a questionnaire. The analysis technique used is Chi Square analysis. The results showed that the influence of the level of knowledge about breast cancer on housewives was mostly in the good category of 60.5%. BSE behavior in housewives is mostly in the category of not doing BSE by 40%. The results of the Chi Square analysis obtained the calculated X2 value of 55.40 with a p value of 0.000 (p <0.05). The close relationship between the level of knowledge of breast cancer with conscious behavior has a strong category indicated by a coefficient of 0.50. The conclusion of this study is that there is a relationship between the level of knowledge of mothers about breast cancer and the BSE behavior.

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Dehydrin (DHN) is known to play an important role in plant response and adaptation to abiotic stresses (drought, high salinity, cold, heat, etc.). Previous research reported the increased expression of DHN in sugarcane stems exposed to drought stress for 15 days which may be controlled by its corresponding stress inducible promoter. The DHN promoter was succesfully isolated from sugarcane variety PSJT 941 (Pr-1DHNSo) and was cloned to pBI121 expression vector fused to a β-glucuronidase (GUS) reporter gene.  The aim of this research was the functional testing of the Pr-1DHNSo promoter through transformation into tobacco plant treated with in vitro drought stress. Genetic transformation of Pr-1DHNSo construct was conducted by Agrobacterium tumefaciens. The transformed tobacco was then subjected to drought stress treatment using 40% PEG 6000  for five sequential incubations (0, 12, 24, 48 and 72 hours). The GUS assay reveal that the transformed tobacco treated with drought stress showed a blue color denoting GUS activity in leaf, stem and root tissues and this expression increased along with the length of the drought treatment. The analysis of gusA gene using real time-qPCR normalized to the L25 reference gene also showed that the expression increased in line with the length of time of drought stress.  The results presented in this study indicated that the Pr-1DHNSo promoter from sugarcane was expressed and induced by drought stress treatment in tobacco.