Yield trials of black rice Cempo Ireng M7 generation of Gamma-Ray irradiation

Cempo Ireng rice is local rice that is rich in anthocyanins. The issue with Cempo Ireng rice is its lengthy harvest life, high habitus, and low production. Plant mutations are one method for overcoming this. The study’s goal is to create better mutant plants with high yields. The study was conducted by placing three lines in plots from December 2019 to August 2020. The results were descriptively evaluated and compared to the control using the T-test. The results showed that the mutant plant was superior to black rice without irradiation (control). The line with the highest productivity value (per hectare) was line 51 which was 8.45 tons/ha. Line 44 has the highest average anthocyanin content of 75.10 ppm.

CRISPR/Cas9-Mediated Targeted Mutagenesis of CYP93E2 Modulates the Triterpene Saponin Biosynthesis in Medicago truncatula

In the Medicago genus, triterpene saponins are a group of bioactive compounds extensively studied for their different biological and pharmaceutical properties. In this work, the CRISPR/Cas9-based approach with two single-site guide RNAs was used in Medicago truncatula (barrel medic) to knock-out the CYP93E2 and CYP72A61 genes, which are responsible for the biosynthesis of soyasapogenol B, the most abundant soyasapogenol in Medicago spp. No transgenic plants carrying mutations in the target CYP72A61 gene were recovered while fifty-two putative CYP93E2 mutant plant lines were obtained following Agrobacterium tumefaciens-mediated transformation. Among these, the fifty-one sequenced plant lines give an editing efficiency of 84%. Sequencing revealed that these lines had various mutation patterns at the target sites. Four T0 mutant plant lines were further selected and examined for their sapogenin content and plant growth performance under greenhouse conditions. The results showed that all tested CYP93E2 knock-out mutants did not produce soyasapogenols in the leaves, stems and roots, and diverted the metabolic flux toward the production of valuable hemolytic sapogenins. No adverse influence was observed on the plant morphological features of CYP93E2 mutants under greenhouse conditions. In addition, differential expression of saponin pathway genes was observed in CYP93E2 mutants in comparison to the control. Our results provide new and interesting insights into the application of CRISPR/Cas9 for metabolic engineering of high-value compounds of plant origin and will be useful to investigate the physiological functions of saponins in planta.

Characterization and Investigation of Stigmasterol Isolated from Rodent Tuber Mutant Plant (Typhonium flagelliforme), Its Molecular Docking as Anticancer on MF-7 Cells

Typhonium flagelliforme is an Indonesian herbal plant used and applied traditionally to treat cancer diseases. Gamma rays have irradiated rodent tuber mutant plant at six doses gray to in-crease the chemical compounds of anticancer activity. The effect of isolated compounds from ro-dent tuber mutant plants has never been studied and published yet. Our study unveiled the poten-tial of stigmasterol as a remarkable cytotoxic agent and the significant contribution of NMR spectroscopy, IR, Mass spectra, QTOF MS towards the isolation and identification of this anti-cancer agent. Stigmasterol was isolated from T. flagelliforme mutant plant. Stigmasterol was more effective against MCF-7 cells with an IC50 value of 0.1623 µM than Cisplatin with IC50 value 13.2 µM. It is the most potential and active fraction in the human breast cancer cell line. The mo-lecular docking study analyzed the chemical profile of stigmasterol to confirm the receptor in agonist binding sites. The prediction of the toxicity of stigmasterol compounds using in silico and analysis of its interaction with the receptor can act as a competitive regulator with a high-affinity binding site on FXR. Stigmasterol has potential as a candidate for an anticancer drug that pro-moting further clinical action.

S-acylation is involved in tonoplast targeting of flax resistance protein M

Several plant resistance proteins require accurate targeting to cellular membranes for effective pathogen defence function. The resistance protein variant M from Linum ussitatissimum (flax) protects plants against the flax rust disease and is specifically targeted to the vacuolar membrane. This localization mechanism involves the S-acylation of cysteine residues within the N-terminus of the protein. Moreover, the M S-acylation and targeting signal likely binds to membranes in the absence of lipid modifications and can switch from the vacuolar membrane to the plasma membrane depending on the S-acylation status. Importantly, plasma membrane targeting was observed when the short targeting signal from M was expressed in an Arabidopsis thaliana pat10 mutant plant. On the other hand, tonoplast localization of the N-terminal S-acylation domain was reconstituted in the mutant plant upon co-expression of two highly related PAT10 enzymes from flax. In contrast to the Golgi and tonoplast targeted Arabidopsis PAT10, the two homologous enzymes from flax mainly localized to the plasma membrane and were partially observed at the vacuolar membrane when expressed in flax as well as in Arabidopsis cells. This implicates that S-acylation of flax proteins could occur at the plasma membrane and that the lipid modification is required for subsequent routing to the vacuolar membrane, which could be also involved in the localization mechanism of M.

Inheritance of a Novel Heterozygous Peanut Mutant, 5-Small Leaflet

ABSTRACT An unusual 5-Small Leaflet mutant plant was found within the ‘Georgia Green' runner-type peanut (Arachis hypogaea L.) cultivar. Subsequent selfing has not established a true-breeding 5-Small Leaflet genotype. It continues to segregate normal and 5-Small Leaflet plants but with a reduced number of normal leaf plants upon selection for 5-Small Leaflet phenotypes after several self-generations. F1, F2, F3, and F4 data suggests that the 5-Small Leaflet trait is dominant or possibly pseudo-dominant. Likewise, the 5-Small Leaflet mutant can only be used as a pollen parent in crosses, and it has approximately a 1:1 ratio of elongated to normal stigmas, respectively, on individual plants. This is an example of a novel heterozygous peanut mutant plant found within the cultivated allotetraploid peanut.

Revolute-Leaf, a New Completely Dominant Mutant in Peanut

ABSTRACT A Revolute-Leaf mutant plant was discovered in an advanced Georgia peanut (Arachis hypogaea L.) breeding line, GA 112702. The mutant had leaf margins that curve downward on each of the four small light-green leaflets. Two cross combinations were used to determine the inheritance of this new mutant. F1, F2, and F3 segregation data strongly supported a single completely dominant gene, designated Rev, controlling the inheritance of the Revolute-Leaf mutant. The F2:3 homozygous revolute-leaf individual plants had shorter mainstem heights, narrower leaflet length and width, narrower canopy width, reduced pod and seed weights, but similar SMK percentages compared to the F2:3 homozygous normal leaf plants resulting from the same closely related cross combination (GA 112702 x Revoluted-Leaf mutant).