Pengaruh Cekaman Aluminium Terhadap Kandungan Asam Organik Dalam Kalus Dan Pinak Tomat (Lycopersicon Esculentum Mill.)

The purpose of this research was to evaluate the effects of Al stress on citric, Malic and oxalic acid content of L. esculentum cv. Intan callus and plantlet, also aluminum content of L. esculentum plantlet. Callus was induced from cotyledone of L. esculentum on Murashige & Skoog (MS) media containing 10-7 M NAA and 10-6 kinetin. The callus was then transferred step wisely at 3 weeks interval to media containing 220, 275, 330, 385, 440, 550, 825, and 1100 μM AlCl3. The callus cultures on the control media and media with the addition of 550 μM AlCl3 were able to regenerate and produce shoots after 8 passages of subculture. The shoots from media with the addition of 550 μM AlCl3 were transferred into the media with addition of 825 μM AlCl3, then to the media with 1100 μM AlCl3. The High Pressure Liquid Chromatography (HPLC) analysis showed that Al stress callus and plantlets contained Malic acid, but no citric and oxalic acid. The content of Malic acid in callus decreased with increasing AlCl3 concentration from 0 to 385 μM. On the other hand, the content of Malic acid in callus increased with increasing AlCl3 concentration from 440 μM to 1100 μM. Similarly, the content of Malic acid in root increased with increasing concentration of AlCl3 from 550 μM to 1100 μM. The result of Neutron Activation Analysis showed that Al content in root decreased as the amount of AlCl3 increased in the media. These results suggested that L. esculentum callus and plantlet respond to the Al stress by producing higher amount of Malic acid

Keragaman Jumlah Salinan Transgen Galur T0 Padi Kultivar Nipponbare Berdasarkan Analisis QPCR Dengan Penanda Gen HptII

The development of transgenic crop using Agrobacterium tumefaciens produces different inserted transgenes, whether copy numbers or location in the plant genome. The research was performed to detect chimeric phenomena based on the transgene quantity analysis in tillers of a clump and of some clumps which were derived from the same calli. CsNitr1–L gene and hptII gene as a marker on a binary plasmid pCAMBIA1300 was transformed into the Nipponbare rice genome using A. tumefaciens strain LBA 4404. Molecular analysis was carried out on three tillers of each four clumps of Nipponbare transgenic T0 generation (events number 1, 2, 3, and 4) and four groups of T0 clump derived from one callus. Three T0 clump samples were collected from each of the four groups of T0 clumps. The results of qPCR analysis showed that the transgene copy numbers of tillers which were derived from one T0 clump were the same. qPCR analysis also discovered that not all plants from one callus demonstrated the same transgene copy numbers. This implies that each T0 rice clump which grows from the transformed calli was needed to be split in the acclimatization step so that the uniform T1 seeds would be obtained

Keragaman Jumlah Salinan Transgen Galur T0 Padi Kultivar Nipponbare Berdasarkan Analisis qPCR dengan Penanda Gen hptII

Keragaman Jumlah Salinan Transgen CsNitr1-L pada Galur Transforman T0 Padi Kultivar Nipponbare. Perakitan tanaman transgenik dengan menggunakan bantuan Agrobacterium tumefaciens menghasilkan penyisipan transgen yang berbeda, baik dalam jumlah salinan maupun letak transgen dalam genom tanaman. Penelitian ini bertujuan untuk menganalisis keberadaan kimera dan jumlah salinan pada tiap anakan dalam satu rumpun dan beberapa rumpun padi Nipponbare transforman T0 yang berasal dari kalus yang sama. Gen CsNitr1-L pada plasmid biner pCAMBIA1300 ditranformasikan ke dalam genom tanaman padi kultivar Nipponbare dengan menggunakan A. tumefaciens strain LBA 4404. Analisis molekuler dilakukan terhadap tiga anakan dari masing-masing empat rumpun tanaman Nipponbare transgenik generasi T0 (event 1, 2, 3, dan 4) dan empat kelompok rumpun tanaman T0 yang berasal dari kalus yang sama. Dari masing-masing kelompok tersebut diambil 3 rumpun T0. Hasil analisis qPCR menunjukkan bahwa anakan-anakan yang berasal dari satu rumpun tanaman T0 memiliki jumlah salinan transgen yang seragam. Selain itu, hasil analisis qPCR juga mengungkap bahwa tidak semua tanaman yang berasal dari kalus yang sama memiliki jumlah salinan transgen yang seragam. Implikasi dari hasil ini bahwa setiap rumpun padi T0 yang tumbuh dari kalus hasil transformasi perlu dipisah saat aklimatisasi supaya benih T1 yang dihasilkan seragam. 

Pengaruh Cekaman Aluminium terhadap Kandungan Asam Organik dalam Kalus dan Pinak Tomat (Lycopersicon esculentum Mill.)

The purpose of this research was to evaluate the effects of Al stress on citric, malic and oxalic acid content of L. esculentum cv. Intan callus and plantlet, also aluminum content of L. esculentum plantlet. Callus was induced from cotyledone of L. esculentum on Murashige & Skoog (MS) media containing 10-7 M NAA and 10-6 kinetin. The callus was then transferred step wisely at 3 weeks interval to media containing 220, 275, 330, 385, 440, 550, 825, and 1100 μM AlCl3. The callus cultures on the control media and media with the addition of 550 μM AlCl3 were able to regenerate and produce shoots after 8 passages of subculture. The shoots from media with the addition of 550 μM AlCl3 were transferred into the media with addition of 825 μM AlCl3, then to the media with 1100 μM AlCl3. The High Pressure Liquid Chromatography (HPLC) analysis showed that Al stress callus and plantlets contained malic acid, but no citric and oxalic acid. The content of malic acid in callus decreased with increasing AlCl3 concentration from 0 to 385 μM. On the other hand, the content of malic acid in callus increased with increasing AlCl3 concentration from 440 μM to 1100 μM. Similarly, the content of malic acid in root increased with increasing concentration of AlCl3 from 550 μM to 1100 μM. The result of Neutron Activation Analysis showed that Al content in root decreased as the amount of AlCl3 increased in the media. These results suggested that L. esculentum callus and plantlet respond to the Al stress by producing higher amount of malic acid

Performance Test of Vegetative Characteristics of Crossed Rice (Oryza sativa L.) Lines of Ciherang Variety X B11143D Line in Telagasari, Karawang Regency, Indonesia

One way to increase the genetic diversity of Ciherang as the superior variety is to cross Ciherang with the donor B11143D line as a New Plant Type (NPT) rice. This study aimed to obtain Ciherang X B11143D lines with the best vegetative characteristics in the field. The research was conducted in rice fields in Talagasari Village, Telagasari District, Karawang Regency, from May to September 2023. This experiment used a single-factor Randomized Block Design (RBD) with three replications of 22 treatments, consisting of 19 Ciherang X B11143D lines and three comparison varieties. The effect of treatment was studied using analysis of variance. The results showed that the rice lines significantly influenced the vegetative characters of Ciherang X B11143D lines in Telagasari, Karawang Regency. Based on the observed characters, several lines were selected i.e. 124.2.3, 94.3.3, and 20.4.4 lines. Those three lines chosen as backcrossing lines performed similarly compared to Ciherang as recurrent parent and inherited several important traits for rising productivity from the B11143D line as donor parent, namely the length and area of flag leaf, the total number of tillers, and the number of productive tillers, which were significantly higher than Ciherang

Resistance Analysis of CRISPR/Cas9 Genome-Edited Chili M2 Mutant Lines against Pepper Yellow Leaf Curl Viral Disease

Pepper yellow leaf curl virus (PepYLCV) infection transmitted by silverleaf whitefly (Bemisia tabaci [Gennadius]) can decrease chili pepper yield up to 100%. At this moment, there is no chili pepper variety resistant to PepYLCV available. Genome editing approach through CRISPR/Cas9 is an effort to develop variety resistance to the viral infection. The purpose of this study was to obtain M2 lines developed by CRISPR/Cas9 system on proliferating cell nuclear antigen (PCNA) gene for resistance to PepYLCV. A total of four M2 lines (C47-7, L84-2, L84-23, and L120-19) consisting of 60 chili plants were tested for their resistance to PepYLCV. PCR analysis was performed to detect the presence (infection) of the virus. The results showed that a total of 35 plants derived from the four lines were resistant to PepYLCV. They consisted of 7 plants from C47-7 line, 11 plants from L84-2 line, 9 plants from L84-23 line, and 8 plants from L120-19 line. PCR analysis confirmed that the resistant plants obtained from this study were negatively infected by the virus. Since not all tested plants were resistant to virus infection, the PCNA gene allele in these resistant lines were most likely heterozigotes. Sequencing of PCNA gene of the resistant lines is needed to confirm that the resistance phenotypes obtained was due to mutation of the gene. Therefore, further selection needs to be performed to obtain stable and PepYLCV-resistant lines

Optimasi Ekstraksi RNA dan Teknik Kloning: Studi Kasus Kloning Gen Heading Date 3a pada Kelapa Sawit

Pembungaan memegang peranan penting bagi tumbuhan karena memfasilitasi rekombinasi genetik, sehingga mendukung perkembangan keragaman genetik yang penting. Keluarga protein phosphatidylethanolamine binding proteins (PEBP) memainkan peran penting dalam mengatur waktu pembungaan dan dormansi benih di beragam spesies tanaman. Penelitian ini bertujuan untuk merancang vektor biner dengan membangun pCAMBIA1300 yang menggabungkan rangkaian gen EgHd3a dari kelapa sawit. Proses konstruksi gen meliputi ekstraksi RNA, sintesis cDNA, amplifikasi gen EgHd3a, kloning gen menjadi vektor kloning, subkloning ke dalam vektor biner pCAMBIA1300, dan diakhiri dengan validasi gen melalui analisis sekuens. Pada ekstraksi RNA, metode PCL-Chisam telah terbukti efektif melalui ekstraksi berulang, meningkatkan kualitas dan kuantitas total RNA. Dalam proses kloaning, metode konvensional menghadapi tantangan dalam memilih lokasi pembelahan yang tepat. Untuk mengatasi kendala ini, penggunaan enzim dengan overhang yang kompatibel diusulkan sebagai solusi potensial. Secara khusus, penggantian BamHI dari BglII telah secara efektif mengatasi tantangan ini. Konfirmasi integrasi fragmen gen ke dalam plasmid pCAMBIA1300 dicapai melalui pengurutan. Meskipun perbedaan diidentifikasi dalam rangkaian EgHd3a-2, perubahan ini tidak berdampak pada asam amino yang dikodekan, sehingga menjaga integritas rangkaian protei

Tracking the morphological diversity of Bucephalandra motleyana Schott (1858 (Araceae) using its commercial name in the proximities of Jakarta, Indonesia

Bucephalandra Schott, Gen. Aroid. (1858) is a genus within Araceae family and assigned to some aquatic plants endemic to Borneo Island, currently representing 31 species. Bucephalandra species are known as ornamental aquatic plants and common for aquascaping. These aquatic plants are highly valued, approximately € 300 in European ornamental aquatic markets and Rp 50,000–700,000 in local markets. We collected 195 specimens of Bucephalandra from 5 ornamental aquatic plant markets in the proximity of Jakarta City, Indonesia. This study is based on repeated confusion with overwhelmed vernacular names assigned for Bucephalandra in the markets. Therefore, the aims of this study are to collect and to identify of Bucephalandra offered in the aquatic plant markets with emphasis on Bucephalandra motleyana Schott 1858. Specimen identification are mostly based on reference specimens stored in the Herbarium Bogoriense Botany Division – Research Centre for Biology – Indonesian Institute of Sciences (LIPI) Cibinong. As result, this study collected 110 specimens belonged to Bucephalandra motleyana Schott 1858 and 85 specimens identified as other species within this genus

Study of aquatic plants and ecological- physics Tempe Lake, Sulawesi Selatan

Aquatic plants are an indicator of the fertility of an aquatic region. The waters of Lake Tempe are the largest waters of the lake area in South Sulawesi. Lake Tempe is located in the western part of Wajo District, precisely in Tempe District, about 7 km from Sengkang City towards the banks of the Walanae River in southern Sulawesi. The area is about 13,000 ha with a maximum depth of 5.5 m and can reach more than 30,000 ha during floods, and during the dry season, the inundation area reaches only 1,000 ha with a maximum depth of 1 m, located above the continental and Australian and Asian plates. This lake is one of the tectonic lakes in Indonesia. Every year silting the lake occurs. The Tempe hydro vegetation and eco-physical research were carried out in October 2017. The purpose of this study was to record aquatic plant species that live in Tempe Lake and observe ecological changes and physical properties of Lake Tempe. Aquatic plants are expected to be able to filter lake water. The results obtained are physical conditions of sharp-smelling water, unpleasant taste, dark brown, and cloudy color. Chemical indicators of NH3-N waters (0.2976-0.0634), PO4-P (0.0172-0.0844) NO2-N (undetectable), NO3-N (1.7131-1.9335), Sulphate (27.761900 - 37.047620), DO (6.88-7.18) and pH (7.88-8.02). There are 14 species of aquatic plants found in these waters. The most dominant species is water hyacinth. In the case of Tempe lake water vegetation results in siltation of the lake area.Keywords: Aquatic plant, Biodiversity, Species, Tempe lake

Introduksi Konstruk Over-Ekspresi Kandidat Gen OsWRKY76 Melalui Agrobacterium Tumefaciens Pada Tanaman Padi Nipponbare

Delivering of Over-Expression Construct OsWRKY76Candidate Gene in Rice cv. Nipponbare throughAgrobacterium tumefaciens. Aniversari Apriana, AtmitriSisharmini, Wening Enggarini, Sudarsono, Nurul.Khumaida, and Kurniawan R. Trijatmiko. Plant geneticimprovement can be done through classical breeding orgenetic engineering. WRKY is a transcription factor involvedin regulating plant defense responses. OsWRKY76 gene islocated in a narrow segment of chromosome 9 which isidentified previously to be related to wide spectrumresistance in rice. A sequence of OsWRKY76 (+1.200 bp)has available in the gene bank and it makes possible toisolate, clone, and construct the gene into over-expressionvector. The aim of this research was to assemble an overexpressionconstruct of OsWRKY76 candidate gene andintroduce it into rice through Agrobacterium-mediatedtransformation. A construct of pCAMBIA-1301::35S::OsWRKY76 has been successfully assembled andtransformed into embryogenic calli of rice cv. Nipponbareusing A. tumefaciens strain Agl-1 and EHA 105. A number of126 independent lines has been produced, in which Agl-1showed 3.8 times more efficient than EHA 105. PCR analysisof randomly selected 25 independent lines showed that allof them positively contained hptII gene, a selectable markerused in the over-expression construct of the OsWRKY76candidate gene. Based on the result, it could be concludedthat the over-expression construct of OsWRKY76 candidategene have been successfully introduced into the tissue ofNipponbare