MORPHOGENETIC EFFECTS OF SEVERAL PLANT GROWTH REGULATORS (PGR) ON IN VITRO DEVELOPMENT OF BINAHONG (Anredera cordifolia L.) LEAF

The aim of this study was to observe the effects of several plant growth regulator on the development of Binahong (Anredera cordifolia L.) leaf in vitro. Binahong is a potential medicinal plant widely used in Indonesia. The leaves used in this research were young leaves collected from the tips of the Binahong vine. The media used was MS medium with several treatments of various plant growth regulators in different concentrations. Both auxins IBA (indole butyric acid) and 2,4-D (2,4- dichlorophenoxyacetic acid) and also cytokinin BAP (benzyl amino purine) were used with concentrations of 0.5-3 ppm added to basic MS media. Fifteen repetitions were done for each treatment. White callus was obtained from MS media added with 1-3 ppm 0f 2,4-D and green callus was obtained from combination of 0.5ppm IBA+0.5ppm BAP, 0.5ppm IBA+1ppm BAP and 1ppm IBA+0.5ppm BAP. A shoot- like callus was induced from the addition of 1 ppm BAP + 0.5 IBA. The growth of roots also occurred on several explants planted on media MS containing both 2,4- and a combination of BAP and IBA. The different result on binahong leaf grown on different treatments of PGR is due to many factors affecting plant morphogenesis in vitro. This in effect can be continued for further research as it can lead to the potential of Binahong as an alternative model plant in tissue culture experiments

Generation and characterisation of a tomato epigenetic recombinant inbred line (epiRIL) population

Heritable changes in gene expression, which are not due to alterations in the DNA sequence, are termed epigenetic modifications and they play an important role in plant development (Pickaard and Scheid, 2014). These changes include those in DNA methylation, chromatin remodelling and levels of small RNAs. This project focuses on the effects of variations in DNA methylation on the cultivated tomato (Solanum lycopersicum). In Arabidopsis, the effects of DNA methylation on plant development have been investigated using mutants in DNA methyltransferase genes. These mutants have also been used to develop epigenetic recombinant inbred line (epiRIL) populations where the genetic background of individuals in the population is identical, but they show variation in genome wide DNA methylation. The aim of this project was to generate and characterize a novel population of tomato epiRILs and identify the effects of the epigenetic variation on the development of tomato. We have established an epiRIL population in a cherry tomato line (WVA 106) where methyltransferase 1 (MET1) gene has been silenced. F2 seeds were generated by colleagues at INRA, (France). Then the epiRIL population, generations F3-F8, were propagated in Nottingham. The population was screened for altered phenotypes. Morphological changes were observed including twisted and fasciated stems, abnormal leaves, exerted style, twisted stamens and low fruit weight. Targeted analysis of the expression of candidate genes underlying the potential phenotypes focused on genes including FW2.2, SlEZ1 and TM8. RNASeq analysis indicated significant changes in gene expression across the genome of selected epiRILs including altered expression of transposons. Analysis using the enzyme McrBC showed that the level of methylation for target genes was lower in the epiRILs than wild type. Changes in DNA methylation in epiRIL lines was also demonstrated using bisulfite conversion and targeted Sanger sequencing and whole genome bisulfite sequencing. The epiRIL population provides a unique resource for investigating the epigenetic basis of trait variation in tomato which could also be applied in other crop species

The effects of 2,4-Dichlorophenoxyacetic acid and leaf surface orientation on callus induction of black betel (Piper betle L. var. nigra)

Black betel (Piper betle L. var. nigra), which has pharmacological properties against a number of infectious disorders, contains secondary metabolites like alkaloids, terpenoids/steroids, flavonoids, and tannins. While preserving plant availability, callus culture can quickly enhance the production of these secondary metabolites. This study aims to determine the combination of the concentration of growth regulator 2,4-Dichlorophenoxyacetic acid (2,4-D) and the optimal position of leaf explants for the induction of black betel callus. This study is an experimental study using a complete randomized design with combinations of treatments between concentrations of 2.4-D growth regulators (0.0 mg/l; 1.5 mg/l; 2.5 mg/l; 3.5 mg/l) and leaf explant position (abaxial contact with media and adaxial contact with media). Quantitative data obtained were analyzed using the Kruskal-Wallis test and Mann-Whitney test on the SPSS program. Meanwhile, qualitative data in callus morphology were analyzed descriptively. The treatment of 1.5 mg/l 2,4-D using abaxial leaf position can be chosen as optimal combination of treatment. The results showed no significant difference (P>0.05) for many of the observed variables between this treatment compared to treatment with higher concentration of 2,4-D. MS media with 1.5 mg/l 2,4-D and abaxial leaf position can induce callus at 2.4 weeks after planting, caused 100 % percentage of explants forming callus, browning score of 2.1, 10% explant contamination, callus growth score of 2.6 and callus morphology in the form of yellowish-white callus color and compact callus texture

Identifikasi Variasi Genetik Berdasarkan Gen D-Loop pada Rusa Timor (Rusa timorensis) dari Jawa Timur dan Bali

Saat ini, populasi rusa timor (Rusa timorensis) dilaporkan cenderung mengalami penurunan, menurunnya populasi rusa timor diakibatkan oleh perburuan liar dan perusakan habitat. Di sisi lain, rusa timor (Rusa timorensis) memiliki potensi yang besar sebagai alternatif sumber protein hewani dan sumber daya genetik di Indonesia. Rusa dimanfaatkan dagingnya sebagai sumber pangan telah banyak dilakukan di berbagai daerah persebaran rusa, Namun sayangnya untuk memenuhi kebutuhan tersebut masyarakat pada umumnya hanya mengandalkan dari hasil berburu dan mengambil dari alam. Maka dari itu, dibutuhkan manajemen konservasi yang tepat untuk menjaga kelestarian rusa timor di habitatnya. Manajemen konservasi yang efektif dapat diupayakan dengan mempelajari latar belakang genetik spesies yang terancam. Penelitian ini dilakukan untuk berkontribusi terhadap penentuan manajemen konservasi yang komprehensif. Penelitian ini bertujuan untuk mengetahui tingkat variasi genetik D-Loop mtDNA pada populasi rusa timor (Rusa timorensis) di kawasan Taman Nasional Bali Barat, Baluran, dan Alas Purwo. Analisis variasi genetik D-Loop (displacment loop) DNA mitokondria ini dilakukan menggunakan sampel noninvasif berupa feses yang diperoleh dari kawasan Taman Nasional Bali Barat, Baluran, dan Alas Purwo. Sebanyak 20 sampel berhasil diekstraksi dan diamplifikasi dengan proses PCR. Sebanyak ±1200 bp berhasil dibaca menggunakan mesin sanger sequencer. Namun hanya 769 bp yang berhasil dibaca jelas dan disejajarkan menggunakan software BioEdit versi 7.2.5 kemudian digunakan untuk proses analisis. Analisis keragaman nukleotida (π), kergaman haplotipe (Hd) menggunakan software DnaSP versi 6.12.03, dan rekonstruksi pohon filogeografi dan estimasi jarak genetik dilakukan menggunakan software MEGA-X versi 10.2.5. Dari 20 sampel yang dianalisis, haplotipe yang berhasil ditemukan sebanyak 11 haplotip dan terdapat 30 situs polimorfisme. Hasil penelitian menunjukkan bahwa variasi genetik D-Loop mtDNA pada populasi rusa timor (Rusa timorensis) di kawasan Taman Nasional Bali Barat, Baluran, dan Alas Purwo masih tergolong tinggi dengan nilai keragaman haplotip (Hd) sebesar 0.88421 dan keragaman nukleotida (π) sebesar 0.00588. Keragaman haplotip (Hd) tertinggi ditemukan di kawasan Taman Nasional Alas Purwo dengan Hd = 0.83333, sedangkan keragaman nukleotida tertinggi ditemukan di kawasan Taman Nasional Baluran dengan π = 0.00950. Hal tersebut mengindikasikan bahwa keragaman genetik masih cukup tinggi dan adanya aliran gen

Anatomical, morphological and physiological leaf characters of black betel (Piper betle L. var. nigra) in varying natural and man-made habitats

Piper betle L. var. nigra or black betel (known as Sirih hitam in Indonesia) contains valuable secondary metabolites, such as alkaloids, flavonoids, saponins, tannins, phenols, carotenoids, steroids and terpenoids. Black betel leaf extract has been shown to have antimicrobial activity thus there is a prospect to be developed as a promising herbal plant. Nevertheless, little information is available about its development as a medicinal plant. This research studies the influence of different habitats on several characters of black betel leaves with the aim to understand the suitable environmental conditions for the optimum growth of black betel plants. We used a survey method and random sampling of black betel leaves in four locations in Java Island, Indonesia, namely Banyuwangi which represents natural habitat, and Karanganyar, Ngaglik and Pakem which represent man-made habitats. Measurements of temperature, humidity, soil moisture and light intensity were carried out at each location. Analyses of leaf area, leaf water content, total leaf chlorophyll content and flavonoid content were undertaken and statistically analyzed using SPSS software. Leaf transverse sections were also observed. The results showed that the environmental parameters differed in the four locations. Leaves samples from the natural habitat in Banyuwangi were significantly different (P<0.05) from the three man-made habitats for chlorophyll and flavonoid content. For water content, significant difference was only for Banyuwangi samples with those from Karanganyar and Pakem. For leaf area, significant difference was only found between the Banyuwangi samples and Karanganyar. Observations on the transverse cross section of midrib of black betel leaves from the four locations showed structures that are generally found in Piper betle species, namely the presence of an epidermal layer, trichomes in the abaxial part of the leaf, several layers of the hypodermis, visible vascular tissue and the presence of secretion cells. There were several differences in the leaf anatomy such as greater number of trichomes on the leaves from Karanganyar, the secretory cells that were more visible in the leaves from Ngaglik and Banyuwangi and the sclerenchymal tissue that was more visible in the leaves from Banyuwangi. Such differences are likely influenced by variations in environmental parameters thus showing that the man-made habitat in the Karanganyar location can affect leaves characters similar to black betel plant grown in its natural habitat in Banyuwangi

Generation and characterisation of a tomato epigenetic recombinant inbred line (epiRIL) population

Heritable changes in gene expression, which are not due to alterations in the DNA sequence, are termed epigenetic modifications and they play an important role in plant development (Pickaard and Scheid, 2014). These changes include those in DNA methylation, chromatin remodelling and levels of small RNAs. This project focuses on the effects of variations in DNA methylation on the cultivated tomato (Solanum lycopersicum). In Arabidopsis, the effects of DNA methylation on plant development have been investigated using mutants in DNA methyltransferase genes. These mutants have also been used to develop epigenetic recombinant inbred line (epiRIL) populations where the genetic background of individuals in the population is identical, but they show variation in genome wide DNA methylation. The aim of this project was to generate and characterize a novel population of tomato epiRILs and identify the effects of the epigenetic variation on the development of tomato. We have established an epiRIL population in a cherry tomato line (WVA 106) where methyltransferase 1 (MET1) gene has been silenced. F2 seeds were generated by colleagues at INRA, (France). Then the epiRIL population, generations F3-F8, were propagated in Nottingham. The population was screened for altered phenotypes. Morphological changes were observed including twisted and fasciated stems, abnormal leaves, exerted style, twisted stamens and low fruit weight. Targeted analysis of the expression of candidate genes underlying the potential phenotypes focused on genes including FW2.2, SlEZ1 and TM8. RNASeq analysis indicated significant changes in gene expression across the genome of selected epiRILs including altered expression of transposons. Analysis using the enzyme McrBC showed that the level of methylation for target genes was lower in the epiRILs than wild type. Changes in DNA methylation in epiRIL lines was also demonstrated using bisulfite conversion and targeted Sanger sequencing and whole genome bisulfite sequencing. The epiRIL population provides a unique resource for investigating the epigenetic basis of trait variation in tomato which could also be applied in other crop species

PENGEMBANGAN BIBIT DURIAN LOKAL DENGAN INDUKSI EMBRIOGENESIS SOMATIK SECARA IN VITRO

Durian memiliki nilai ekonomis tinggi baik sebagai salah satu komoditas hortikultura yaitu buah untuk konsumsi di pasar lokal maupun internasional. Salah satu hambatan adalah persediaan bibit yang tidak mencukupi dan metode perbanyakan yang merusak pohon induk dengan karakter unggul. Penelitian ini bertujuan untuk menghasilkan embrio somatic dari daun durian secara in vitro. Penelitian dilakukan di Laboratorium Kultur Jaringan dan Kebun Percobaan FMIPA, UNY. Metode yang digunakan dalam penelitian ini adalah induksi menggunakan zat pengatur tumbuh : 2,4_D, BAP, IBA dan NAA dalam media MS. Hasil penelitian menunjukkan penambahan 3 ppm 2,4-D dalam media MS mampu memicu kalus embriogenik dari daun durian muda yang sudah membuka. Induksi embrio dari kalus belum berhasil dilakukan dengan pemindahan pada media dengan konsentrasi dan jenis ZPT yang sama dan berbeda. Perlu dilakukan lagi eksplorasi dalam mencari media tepat untuk memunculkan mebrio dari kalus embriogenik

Agrowisata belimbing Karangsari, potensi lokal Kota Blitar sebagai sumber belajar biologi untuk meningkatkan literasi sains peserta didik

The utilization of local potential as a biology learning source is still not maximized. This research focuses on integrating the local potential of Blitar, namely Karangsari’s Starfruit Agrotourism which was developed into a student worksheet app to increase students' science literacy. Karangsari’s Starfruit Agrotourism can be used as a learning resource on biodiversity material by discussing the diversity of starfruit (Averrhoa carambola) varieties, the use of starfruit, and efforts to preserve starfruit. Utilizing local potential and implementing the Sustainable Development Goals (SDG) in biology learning aligns with the Merdeka Belajar curriculum. This study used the ADDIE model which consists of analysis, design, development, implementation, and evaluation stages. The final product of this development research is a student worksheet app implemented to enhance science literacy in class X of students at SMAN 1 Blitar. Data was collected with needs questionnaires, teacher interviews, assessment questionnaires for material experts, media experts, biology teachers, student response questionnaires, and science literacy skills tests. The data obtained were statistically analyzed tests between subjects. Research results concluded as Karangsari’s starfruit agrotourism applied as the materials of the student worksheet app was declared feasible by experts, there are differences in science literacy between students who had taken part in learning while using the worksheet and students who did not, and the effect/contribution given by student worksheet apps to science literacy is 58%.Abstrak: Pemanfaatan potensi lokal sebagai sumber belajar biologi masih belum maksimal. Penelitian ini berfokus pada pemanfaatan potensi lokal Kota Blitar yaitu Agrowisata Belimbing Karangsari yang dikembangkan menjadi E-LKPD untuk meningkatkan literasi sains siswa. Agrowisata Belimbing Karangsari dapat dijadikan sebagai sumber belajar materi keanekaragaman hayati dengan membahas keanekaragaman varietas belimbing manis (Averrhoa carambola), pemanfaatan belimbing, dan upaya pelestarian belimbing. Pemanfaatan potensi lokal dan implementasi Sustainable Development Goals (SDG) dalam pembelajaran biologi sejalan dengan kurikulum Merdeka Belajar. Penelitian ini menggunakan model ADDIE menurut Aldoobie (2015) yang terdiri dari tahapan analisis, desain, pengembangan, implementasi, dan evaluasi. Produk akhir dari penelitian pengembangan ini adalah E-LKPD yang diimplementasikan untuk meningkatkan literasi sains pada siswa kelas X di SMAN 1 Blitar. Pengumpulan data dilakukan dengan angket kebutuhan, wawancara guru, angket penilaian ahli materi, ahli media, guru biologi, angket respon siswa, dan tes keterampilan literasi sains. Data yang diperoleh dianalisis secara statistik dengan uji test between subject. Hasil penelitian menyimpulkan bahwa (1) agrowisata belimbing karangsari yang diterapkan pada E-LKPD dinyatakan layak oleh ahli, 2) terdapat perbedaan literasi sains antara siswa yang pernah mengikuti pembelajaran dengan menggunakan E-LKPD dan siswa yang tidak; (3) pengaruh/kontribusi yang diberikan E-LKPD terhadap literasi sains sebesar 58%

APPLICATON OF MYCORRIZA ON PLANTING MEDIA OF TWO TOMATO VARIETIES TO INCREASEVEGETABLE PRODUCTIVITY IN DROUGHT CONDITION

A method used for the development of dry areas/marginal lands is the improvement of soil structure and addition to the media to support the growth and development of crops. Tomatoes have the potential to be developed in marginal lands due to its high nutrition, high in demand and easy to be cultivated. One of the method used to improve planting media is the addition of microorganism such as mycorrhiza which can help the absorbtion of water and nutrition for plants. The interval of irrigation is used as a simulation of drought. This research aim was to observe the effect of mycorrhiza in the soil on the growth and development of tomato with several treatments of irrigation. The method used was the addition of 4 g of mycorrhiza  per polybag (size 30x30 cm2), using Complete Randomized Design. There were 6 combinations of treatments. The treatments were : 3 interval of irrigation (every day, every 7 days and every 14 days), and 2 treatments of mycorrhiza (0 g and 4 g). There were 3 repetition for each combination of treatments. The results showed that the addition of mycorrhiza can increase significantly plant fresh and dry weight and also root length. The difference in plant height, number of branches, number of leaves, plant growth rate and percentage of infection were caused by the difference in irrigation interval. The difference in the varieties used also contribute to a difference in the percentage of infection. Further research must be made on the effect of mycorrhiza with addition of inorganic fertilizer to increase the growth and development of tomato plants in water stressed condition. Keywords:   mycorriza, tomato, draught simulatio