Obtaining of transgenic potato (Solanum tuberosum L.) cultivar IPB CP3 containing LYZ‐C gene resistant to bacterial wilt disease

Bacterial wilt caused by Ralstonia solanacearum is one of the most important bacterial diseases in potato production. This study aimed to obtain the transgenic potato (Solanum tuberosum L.) cultivar IPB CP3, containing LYZ‐C gene encoding for lysozyme type C, resistant to bacterial disease caused by R. solanacearum. Genetic transformation using Agrobacterium tumefaciens LBA4404 to 124 internode explants resulted in the transformation efficiency of about 47.58% with a regeneration efficiency of approximately 30.51%. Gene integration analysis showed that 16 clones were confirmed as transgenic clones containing the LYZ‐C gene. Analysis of resistance to R. solanacearum of three transgenic clones showed that all three transgenic clones were more resistant than a non‐transgenic one. This result showed that the LYZ‐C gene integrated in the genome of transgenic potato increased the resistance of potato plants to R. solanacearum. We obtained two transgenic clones considered resistant to bacterial wilt disease

Peranan Pseudomonas aeruginosa ATCC 15442 terhadap Pertumbuhan Tanaman Cabai Rawit (Capsicum frutescens L.) Varietas Bonita pada Kondisi Salin: The Role of Pseudomonas aeruginosa ATCC 15442 on the Growth of Bonita Hot Pepper (Capsicum frutescens L.) Variety in Saline Conditions

Hot pepper (Capsicum frutescens L.) is one of the food crop commodities that is widely cultivated in Indonesia. Salinity stress can reduce the bioavailability of potassium and its uptake by plants, which will ultimately reduce plant growth and production. One way to reduce the effect of salinity and increase potassium uptake by plants is to use potassium-solubilizing bacteria. One of the bacteria that can solubilize potassium is Pseudomonas aeruginosa. This research aims to study the role of Pseudomonas aeruginosa ATCC 15442 on the growth of the Bonita variety of hot pepper under saline conditions. This study used a completely randomized factorial design with four concentration levels of NaCl treatment: 0, 4, 8, and 12 g/L, as well as two levels of bacteria administration: without bacteria and with bacteria. The results showed that the application of bacteria, salt concentration, and the interaction of the two had no effect (p-value> 0.05) on the growth of hot pepper plants with the observed parameters namely plant height, number of leaves, shoot fresh weight, root fresh weight, shoot dry weight, root dry weight, and chlorophyll content

Alkaline Phosphatase Activity of Plant Growth-Promoting Actinomycetes and Their Genetic Diversity Based on the phoD Gene

Actinomycete is one of the beneficial bacteria groups inhabiting rhizosphere soil. They can promote plant growth through various mechanisms. In the previous study we have isolated rhizosphere actinomycetes from maize rhizosphere with direct plant growth promotion characters. The aims of the present study were to analyze the ability of maize rhizosphere actinomycetes to solubilize phosphate, determine alkaline phosphatase activity, and study their genetic diversity based on phoD gene. Thirteen rhizosphere actinomycete isolates were able to solubilize phosphate at concertation range 55.84±2.27 mg/L to 144.48±5.71 mg/L. The activity of extracellular alkaline phosphatase was exhibited by all maize rhizosphere actinomycetes isolates in various level ranging from 0.08 mU/mL to 0.51 mU/mL. The phoD gene, one of the three homologous genes which encode alkaline phosphatases, was successfully detected in all isolates and identified as alkaline phosphatase D of Streptomyces spp. The partial phoD sequences of the isolates were located within metallophosphatase domain of alkaline phosphatase D. Alignment analysis showed that the deduced amino acid sequences of PhoD were mostly conserved in the isolates and Streptomyces spp. Essential residues involved in the active core arrangement of PhoD which binds metal ion cofactors were conserved. Constructed phylogenetic tree showed that the isolates were divided into two groups within PhoD cluster. PhoD of the isolates and Streptomyces spp. had closer relationship to purple acid phosphatase compared to other homologous PhoA and PhoX which form separate cluster. Generated three-dimensional structure model of partial PhoD had high similarity to alkaline phosphatase D of Bacillus subtilis (2YEQ) and showed overlapping structure based on super-positioning analysis

Construction of RNA Interference Vector to Silence Aluminum Tolerance Gene Candidate in Rice cv Hawara Bunar

One of the aluminum (Al) tolerance gene candidates, namely B11 gene, has been successfully isolated from Al-tolerant rice cv Hawara Bunar. However, the role of the gene in Al tolerance in rice has not been known. RNA interference (RNAi) technique is an effective tool to examine the biological function of the target gene in plant. The objective of the research was to construct RNAi recombinant vector carrying untranslated region of the B11 gene. RNAi recombinant vector carrying 195 bp sized 3′UTR_B11 fragment as a double-stranded RNA (dsRNA) trigger has been successfully constructed using GATEWAY™ cloning technology, pENTR™/D-TOPO® as a shuttle vector, and pANDA vector as a destination vector. RNAi construct was successfully introduced into Agrobacterium tumefaciens AgL0, and has been infected to rice cv Hawara Bunar. Analysis of putative transgenic rice showed eight of 20 plants were transgenic carrying the B11-RNAi construct

Establishment of transgenic potato cultivar IPB CP1 plants containing gene encoding for superoxide dismutase to increase the abiotic stress tolerance

Potato ( Solanum tuberosum L.) cultivar IPB CP1 is suitable as a raw material for the potato chip industry. Potato plants are sensitive to various abiotic stresses such as drought, aluminium and salinity, which induce reactive oxygen species (ROS). ROS is very toxic to plant cells. Superoxide dismutase (SOD) is one of the enzymes that catalyse ROS to H2O2 and O2. This study aimed to establish transgenic potato cv. IPB CP1 plants containing the MmCuZn‐SOD gene that are tolerant to various abiotic stresses. Genetic transformation using internodes without buds as explants produced putative transgenic potato with a transformation efficiency of 51.25% and a regeneration efficiency of 38.87%. Integration analysis of the MmCuZn‐SOD transgene in putative transgenic plants by polymerase chain reaction (PCR) with a set of specific primers showed that eight plants contained the MmCuZn‐SOD gene under the control of the 35S CaMV promoter. In vitro salinity stress, aluminium stress, and drought stress assays showed that transgenic plants had a higher number of roots and total root length than non‐transgenic ones. These results indicate that transgenic potato cv. IPB CP1 plants are more tolerant to abiotic stresses than non‐transgenic ones

Identifikasi Marka Gen Ketahanan Hawar Daun Bakteri Pada Galur Padi Introduksi Dan Galur Dihaploid

Identification marker of bacterial leaf blight (BLB) gene resistance on introducted and dihaploid rice germplasm. Bacterial leaf blight (BLB), caused by bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo), is one of the most devastating diseases in rice. The use of BLB-resistant rice varieties is one of the most efficient ways to protect rice from this disease. BLB-resistant varieties can be produced through the breeding program by using the diverse rice germplasm. The objective of this research was to identify BLB resistance gene on 37 introducted and dihaploid rice lines derived from wide genetic background double crossing from local rice : IR54/Parekaligolara and Bio110/Markuti, by using the molekular markers. As control plant used 23 differential varieties (monogenic lines/IRBB) and TN1 (susceptible). All plants tested were inoculated by 3 selected dominant BLB races (Race III, IV dan VIII). The selected of 19 polymorphism molecular markers used to identify the BLB resistance genes on rice lines tested. The result of this research showed that 4 rice lines were resistance to all BLB races tested. Three molecular markers were specific associated with resistant gene to Race III (Xa7-STS40, Xa1-STS14 and Xa4-STS50); Race IV (Xa1-STS5, Xa4-STS50 and Xa26-STS1), and Race VIII (Xa21-STS6, Xa7-RM20590 and Xa7-STS40). These markers could be utilized for the selection process the developmen of BLB resistance rice lines breeding program

Manipulasi Nutrien dan Zat Pengatur Tumbuh untuk Meningkatkan Produksi dan Kualitas Buah Stroberi (Fragaria x ananassa) Kultivar BAT-1

Stroberi kultivar BAT-1 semakin jarang dibudidayakan oleh petani Indonesia karena rasa buahnya kurang manis, padahal kultivar tersebut mampu beradaptasi dengan kondisi lingkungan tropis. Hal tersebut disebabkan perbaikan kualitas dan produktivitas stroberi adaptif itu masih sulit dilakukan dengan praktik pertanian biasa. Tujuan penelitian ini adalah untuk meningkatkan produksi dan kualitas buah stroberi BAT-1, yaitu ukuran, warna, rasa manis dan umur simpan, dengan memodifikasi nutrien dan manipulasi zat pengatur tumbuh (ZPT). Penelitian ini dilaksanakan pada Agustus – Desember 2021 (musim hujan) dan Februari – Juni 2022 (musim kemarau), di kaki Gunung Halimun-Salak, Kabupaten Bogor. Penelitian menggunakan rancangan acak lengkap faktorial: Perlakuan tahap pertama berupa pemberian kalium dan kalsium, dan diukur parameter vegetatifnya. Untuk tahap kedua diberikan perlakuan asam indol butarat dan brassinolida, dan diukur parameter generatifnya. Hasilnya, perlakuan kalium dan kalsium mempercepat dan mempersingkat rentang waktu berproduksi tanaman; dan modifikasi nutrien yang berkombinasi dengan ZPT mampu meningkatkan bobot buah, kandungan antosianin buah, nilai kemanisan serta menurunkan nilai total asam. D3I3B2 (PPC + CaCl2 2 g L-1 pada daun dan ZPT IBA 3 mM + BL 0.2 µM pada buah) merupakan perlakuan terbaik, di kedua musim tanam. Brassinolida (0.2 µM) mampu memperlama daya simpan buah setelah panen.-1Kata kunci: Brassinolide, brix, IBA, kalium, kalsium.Indonesian farmers less frequently cultivate strawberry cv.BAT-1 because the fruit is less sweet, even though the cultivar is more adaptable to tropical conditions. Meanwhile, improving the quality and productivity of this cultivar is difficult for ordinary farmers. This study aimed to help increase the production and quality of strawberry BAT-1, i.e., the size, color, fruit sweetness, and shelf life, by manipulating some nutrients and hormones. This research was conducted in August - December 2021 and February - June 2022 (for the rainy and dry seasons, respectively) in Halimun-Salak, Bogor. This study used a factorial complete randomized design. The first stage of treatment was giving potassium and calcium and measuring vegetative parameters. Treatment with IBA and brassinolide was applied for the second stage, and generative parameters were measured. The results were that the treatments of potassium and calcium advanced and shortened the productive period. Combined with two growth regulators, they increased the fruit weight, anthocyanin content, and sweetness and decreased total acid value. D3I3B2 (PPC + CaCl2 2 g L on leaves and IBA 3 mM + BL 0.2 µM on fruits) was the best treatment in both seasons. Brassinolide (0.2 µM) extended the shelf-life of the fruit. Keywords: Brassinolide, brix, calcium, IBA, potassiu

Pertumbuhan Planlet Lidah Mertua (Sansevieria sp.) Blue Leaf dari Kultur Kalus

Lidah mertua (Sansevieria sp.) Blue Leaf is a slow growing ornamental plant. It is also difficult to reproduce. Therefore market demands for this plant is impossible to fulfill through conventional methods of propagation such as leaf cutting or layering. Tissue culture is one of the alternatives to solve the problem of plant propagation. In vitro culture of Sansevieria calli was induced for shooting, and rooting afterwards. The basic media of WP was superior than MS in producing multiple shoots, and this WP media containing of BAP 5 mg/L and NAA 0,5 mg/L (WH1) gave the highest number of shoots and leaves on plantlets, as well as the best growth performance. The shoots started to appear two weeks after culture. Plantlets from WH1 rooted in higher frequency as well on rooting media. A mixture of compost, sand, and zeolit (4:4:1) enriched with a nutritive solution, composting of 1⁄4 WP plus IBA 0.5 mg/L, was a poreous media that was able to induce rooting of this plant.&nbsp

Analisis Metabolomik pada Interaksi Padi dan Bakteri: Metabolomics Analysis on the Interaction of Rice and Bacteria

Mutualistic plant-microbe interaction can increase the availability or absorption of nutrients and affect plant metabolism. Metabolomics is the study of all metabolites contained in living beings at a specific time. This research aims to study the effect of metanotrophic bacteria (BGM 9 isolate) and nitrogen-fixing bacteria (ITJ 7 isolate) on rice plant growth and reveal the information about metabolites produced by rice plant interacting with the bacteria. Three rice plant varieties (Ciherang, Hawara Bunar, and IR64) were grown on liquid media (Yoshida solution) and were inoculated with the bacteria five weeks after planting. Metabolites in the liquid media were detected seven days after inoculation using Liquid Chromatography-Mass Spectrometry and analyzed using MassLynx v4.1. The results showed that the bacterial inoculation had significant effect on rice growth. Chromatograms resulted from LC-MS showed there are 34 unique peaks from Ciherang, 21 unique peaks from Hawara Bunar, and 30 unique peaks from IR64. The unique peaks in each variety were affected by the bacteria inoculation treatments.&nbsp

Horizontal Gene Transfer and Population of Phyllosphere Bacteria on Transgenic and Nontransgenic Cotton

The possibility of horizontal gene transfer of plant genomic DNA and bacteria in the soil, particularly as this relates to the possible transfer of genes encoding antibiotic resistance, has been seen as hazard associated with genetically engineered plants. It is hypothesized that introduction of bacterial genes into the plant genome leads to a higher probability of gene transfer from plants to bacteria due to the presence of homologous sequences. Bollgard (BG) cotton was constructed through the introduction of cry1A(c) gene, encodes for insecticidal activity againts Lepidopteran pests, together with genes for spectinomycin/streptomycin resistant (aad) and kanamycin resistant (nptII), into the genome of a conventional cotton variety, Delta Pine (DP). The aim of this study were to evaluate the ability of naturally competent Acinetobacter calcoaceticus strain ADP1 to take up and integrate transgenic plant DNA based on homologous recombination under optimized laboratory condition, and to compare phyllosphere microbial population resistant to antibiotic on leaves of transgenic and nontransgenic plant. The results showed that transformation of ADP1 cells with Bollgard DNA was not detected on nitrocellulose membrane nor in sterile soil. Total phyllosphere bacterial population on leaves collected from one month after planting were 1.3 x 108 and 1.6 x 108 cfu/g leave fresh weight for BG and DP, respectively. Samples collected after three month contained 5.9 x 107 and 7.1 x 107 cfu/g leave fresh weight for BG and DP, respectively. This study also showed that there was no significant difference of phyllosphere bacterial population resistant to streptomycin and kanamycin on leaves of BG or DP samples collected from one or three month after planting