<div>Background: Pigmented rice particularly, red and black rice have come into prominence due to their promising antioxidant property. In Assam, all major ecotypes of paddy viz., Ahu, Sali, Bao, Boro have large number of indigenous landraces with red kernel. Although Assam is recognised as a secondary centre of origin and diversity yet gene erosion is assuming serious proportion. It is acknowledged that many indigenous cultivars are facing extinction and there is apprehension that many may be lost even before they are evaluated and characterised. Among different methods, seed protein profile is recognised as fingerprint for a race by International Seed Testing Association (ISTA) as it can resolve interspecific and intraspecific variation without ambiguity.&nbsp; 
 
Methods: The present study involves, molecular characterisation and genetic diversity analysis of pigmented and non-pigmented landraces. Seed protein profiles for the paddy landraces were worked out by SDS-PAGE. SDS-PAGE was carried out with two phase gradient buffer system with upper stacking gel and lower separating gel to ensure denaturation of protein while migrating through it. 
 
Result: Among the twenty landraces a total of 15 proteins bands were observed in the size range of 12.1 Kd to &gt;122.0 Kd. Highest protein bands were observed in cultivar Ranjit Amon, Bismuthi and Boro (ecotype) all with 15 protein bands. On the other hand, lowest protein bands were recorded in Amona with only 9 bands. Among the protein bands six with molecular weight &gt;122.0 Kd; 112.0 Kd; 97.4 Kd, 90.0 Kd and 15.0 Kd were consistently found in all the landraces. The dendrogram revealed a total of four clusters. The landrace Amona (red) did not belong to the any cluster and most distantly related to others. On the other hand, some are genetically identical. Agnisali, Suhagmani and Tengeri (all white) had no genetic distance between them and hence identical at molecular level. Within the same cluster both red and white rice landraces were observed. Our study suggested that several proteins bands were detected only on specific rice and it could be as biochemical markers for further research.</div>

Genetic Diversity of Seed Storage Proteins in Pigmented and Non-pigmented Indigenous Paddy (Oryza sativa L.) Landraces of Assam, India

<div>Background:&nbsp;Selecting an appropriate extraction technique and optimization of extraction conditions must be done to improve the efficiency and productivity of natural colorant. 
 
Methods: Flesh and peels of four different raw materials viz., beetroot, carrot, pomegranate and grape were extracted independently using 100% ethyl alcohol using cold solvent extraction method adopting 1: 1 and 1: 2 w/v solid to solvent ratio for 24 and 48 hrs, forming 8 treatments replicated thrice in three factorial CRD. The collected extracts were evaporated at 60oC; per cent yield and pigment content were calculated. 
 
Result: Extract yield and pigment content were significantly influenced by raw material, solid to solvent ratio and duration of extraction. Extract yield was higher in flesh of carrot and beetroot as well as in peels of grape and pomegranate. Anthocyanin content in grape and pomegranate and betalain content in beetroot were high in peel where as &beta;-carotene content was higher in carrot flesh extract. Per cent yield and pigment content increased with increased duration of extraction and solid to solvent ratio. Requirement of more solvent and longer extraction time are the drawback of conventional extraction methods and there is a growing demand for non-conventional techniques to enhance yield and maintain pigment stability.</div>

​Standardization of Cold Solvent Extraction Parameters for Natural Red Pigments

<div>Chloroplasts are involved in many cellular processes and play an important role in plant stress response. In this study, the effect of drought on different physiological characteristics and the isolation of chloroplasts from leaves of drought-stressed and control plants of common bean (Phaseolus vulgaris L.) are reported. Two cultivars differing in the response to drought were analysed - Tiber and more sensitive Starozagorski &egrave;ern. The results of relative water content showed that plants were under mild stress conditions after six days without watering. Negative effects of drought stress on the photosynthetic rate and chlorophyll fluorescence parameters of both cultivars were observed. The yield of isolated chloroplasts in control samples of both cultivars was at least three times higher compared to drought-stressed samples. The intactness of the isolated chloroplasts was checked and used for protein isolation. The results of the study suggested that even mild drought stress strongly affected the isolation of intact chloroplasts. Still, intact chloroplasts were enriched and were suitable for downstream proteomic analysis.</div>

<div>Impact of drought stress on physiological characteristics and isolation of chloroplasts in common bean (Phaseolus vulgaris L.)</div>

<div>Background: Biofloc technology is a minimal or zero water exchange technology which exerts beneficial effects on water quality, improves feed conversion ratio by recycling microbial protein in the culture of commercially important finfishes and shellfishes. This culture technique also improves the pigmentation in the ornamental fish culture. The presence of pigment producing bacteria and the absorption of the pigments by the ornamental fishes under biofloc technology would pave a long way to boost the production and export market of the high value fishes. Therefore, the present study evaluated the use of bacterial pigments isolated from the biofloc culture in the diets of Swordtail ornamental fish for its colouration and pigment absorption ability. 
 
Methods: A 30 days trial was conducted to evaluate the colour enhancement in swordtail through the supplementation of pigment produced from the bacteria in their diets. Swordtail fishes (1.2 &plusmn; 0.01 g) were stocked in 100 litres tanks (20 nos / tank) in triplicates. Fish fed with normal diet served as control and diet supplemented with pigment (50 mg/kg) isolated from the bacteria Exiguobacterium profundum (T1), Chryseobacterium joostei (T2), Staphylococcus pasteuri (T3), Staphylococcus arlettae (T4) served as treatments. The characteristic features of the pigments isolated from the four different bacteria were checked and showed enhanced antibacterial, total antioxidant activity and the reductive ability. 
 
Result: Significant difference was observed in water quality parameters except temperature between the experimental groups. Growth parameters showed significant difference between control and treatment. Highest carotenoid concentration was found in fishes fed with pigments extracted from Staphylococcus pasteuri compared to other experimental groups. The present study proved the incorporation of bacterial pigments in the diets of Swordtail enhanced the total carotenoid concentration.</div>

<div>Characterisation of Pigment Producing Bacteria Isolated from Bio Floc Ponds and Its Colour Enhancement Effect in Xiphophorus helleri</div>

Background: Astragalus is one of the largest angiosperms and has important economic value. The chloroplast genomes (Cp) of most plants in its family have been sequenced and annotated, but there are fewer studies on the characteristics of the chloroplast genomes and codon usage bias of Astragalus. In this study, we sequenced and annotated the complete chloroplast genomes of three Astragalus species, that systematically compared the chloroplast genome and codon usage characteristics of three Astragalus. 
 
Methods: In this study, we used three Astragalus species as materials. Firstly, we sequenced and assembled their chloroplast genomes. Subsequently, we analyzed the codon usage bias of the chloroplast genomes of the three medicinal Astragalus species using software such as CodonW, CUSP and SPSS. 
 
Result: The results illustrated that the length of Astragalus chloroplast genome ranges from 122,815 bp (A. dahuricus) to 123,729 bp (A. melilotoides Pall.). There were 107-110 genes, including 75-76 protein-coding genes (PCGs), four ribosomal RNA genes (rRNAs) and 28-30 transfer RNA genes (tRNAs). In addition, the results illustrated that codons in three Cp genomes from Astragalus had the same 11 Optimal codons and ending with A/U. The codon usage frequency of five model creatures were compared and the results revealed that the codon preferences of Astragalus and five model creatures were pretty significant, the natural selection was the main factor in codon preference. Our research provides some value for the researches of chloroplast genetic engineering and molecular breeding in Astragalus.

Comparison of the Complete Chloroplast Genomes in the Astragalus

Fourteen pigmented hill rice cultivars along with a non-pigmented one were studied for ascertaining the extent of their nutritional and genetic diversity. Moisture contents of the fourteen pigmented hill rice cultivars ranged from 7.49 to 10.10 % along w.ith the contents (% dry weight) of nutrients viz., crude fat (4.37 &ndash; 5.27) and crude protein (9.27 - 11.42). The corresponding range of values for the contents (mg/100g dry weight) of microminerals Zn and Fe were form 3.42 to 4.28 and from 3.21 to 4.18 respectively. The pigmented germplasm had anthocyanin content (mg cyanidine 3-O glucoside /100g) ranging from 3.58 to 7.86, total phenolic content (mg GAE/100 g)from 67.89 to 89.43, flavonoid content (mg QE/100g)from 57.75 to 78.74 and antioxidant activity from 19.56 to 29.29 %. Analysis of variance indicated that the varietal effect on each of the parameters studied was highly significant. There was significant correlation amongst the different parameters of the rice genotypes analysed. Results of the biochemical screening of these cultivars may lead to selection of promising candidates that can be used for developing improved lines in future breeding programmes. 
&nbsp;

Assessment of nutritive and antioxidant properties of some indigenous pigmented hill rice (Oryza sativa L.) &nbsp;cultivars of Assam

<div>Background: Legumes and cereals microgreens are consumed both in healthy nutrition and as a natural food against many degenerative diseases due to their rich bioactive content. The research was carried out to determine the biochemical contents of 5 legumes and 7 cereals. 
 
Methods: The research was conducted in Van Yuzuncu Yıl University Field Crops Department Laboratory in 2020, in a controlled growing environment, according to the completely randomized experimental design. Total antioxidant activity, total phenolic, total flavonoid and total ascorbic acid contents and total Chlorophyll, Chlorophyll a, Chlorophyll b and Carotenoid contents and correlation between them were determined. 
 
Result: The differences between the means of all traits were found to be significant. The biochemical contents of cereals were 1.6-7 times higher than legumes. There was no big difference between the pigment values. In legumes, Bilensoy alfalfa had the highest total antioxidat activity and total flavonoid content, Uzbek lentil total phenolic content and Goynuk bean had the highest Total ascorbic acid. In cereals, Kirklar oat had the highest total antioxidat activity, total phenolic, total flavonoid and total ascorbic acid&nbsp; contents, while Larende barley had the lowest values (except total ascorbic acid content). While total Chlorophyll and Chlorophyll a amounts were highest in Kirklar barley and lowest in Uzbek lentils, the opposite situation occurred in Chlorophyll b and Carotenoid amounts. There was no significant correlation between biochemical parameters in legumes. There was a positive and significant correlation between total phenolic content with total antioxidant activity and between total ascorbic acid with total fenolic content in cereals. There were positive and significant correlations between all pigment parameters in both legumes and cereals.</div>

Determination of Biochemical Composition and Pigment Content in Legume and Cereal Microgreens

Background: Euchresta tubulosa is a medicinal plant in the Fabaceae family, traditionally used in Chinese herbal medicine for treating ailments such as sore throat and esophageal cancer. Despite its pharmacological importance and endangered status, no complete chloroplast genome data had been reported for this species or its genus prior to this study. Chloroplast genomes, being highly conserved and maternally inherited, serve as powerful tools for species identification, phylogenetic inference and genetic diversity assessment in plants. 
 
Methods: Fresh leaves of E. tubulosa were collected and genomic DNA was extracted using the cetyltrimethylammonium bromide (CTAB) method. High-throughput sequencing was carried out on the Illumina NovaSeq6000 platform. Clean reads were assembled into a complete chloroplast genome using NovoPlasty, with a related species as the seed reference. Genome annotation was performed using DOGMA and validated manually. Codon usage patterns were analyzed using RSCU values and simple sequence repeats (SSRs) were identified with MISA software. A maximum likelihood phylogenetic tree was constructed using PhyML, based on complete chloroplast genome sequences from 14 related species. 
 
Result: The assembled chloroplast genome of E. tubulosa measured 153,960 bp and exhibited a typical quadripartite structure, comprising a large single-copy region (LSC), a small single-copy region (SSC) and two inverted repeats (IRs). The overall GC content was 36.3%. A total of 127 genes were identified, including 44 related to photosynthesis, 72 involved in self-replication, 5 with other biological roles and 6 hypothetical or unknown genes. Codon usage analysis revealed 31 codons with RSCU &gt; 1, showing a strong bias toward A/U-ending codons. A total of 109 SSR loci were detected, the majority of which were located in intergenic spacer regions. Phylogenetic analysis clearly positioned E. tubulosa as an independent lineage within the Fabaceae, supported by high bootstrap values. The genomic information provided by this study lays a foundation for future research in molecular taxonomy, phylogenetics and conservation of this endangered medicinal plant.

Features and Phylogenetic Analysis of the Chloroplast Genome of the Medicinal Plant Euchresta tubulosa

Background:&nbsp;Entada phaseoloides&nbsp;(L.) Merr (Fabaceae)&nbsp;is a large woody climber that is found widely in southern China and other tropical and subtropical areas worldwide. The genus Entada&nbsp;contains ~30 species, and E. phaseoloides&nbsp;is most commonly found in China. The E. phaneroneura&nbsp;and E. pervillei&nbsp;are endangered species. Previous studies had focused on medicinal components, transcriptional regulation, and nuclear genomes. The chloroplast genome of Entada&nbsp;has not been reported, and little is known about the phylogenetic relationships within the Entada. In this study, we performed short-read sequencing of E. phaseoloides&nbsp;and assembled and analyzed its chloroplast genome.

Methods:&nbsp;Dry specimen leaves of E. phaseoloides&nbsp;were subjected to DNA extraction and sequenced using the Illumina Novoseq platform. The chloroplast genome was assembled using Get Organelle, annotated using CPGAVAS2 and Geneious Prime. Repeat sequences and SSR analysis were performed using the Reputer and MISA programs, respectively.&nbsp;Phylogenetic analyses were performed using IQTREE and MrBayes software.

Result:&nbsp;The complete chloroplast genome of E. phaseoloides&nbsp;is 159,963 bp in length and has a quadripartite structure with large single copy of 89,972 bp and a small single copy of 19,309 bp separated by inverted repeats of 25,341 bp. A total of 112 genes in E. phaseoloides&nbsp;comprised 78 protein-coding genes, 30 transfer RNA genes, and 4 ribosomal RNA genes. The distribution of simple sequence repeats and long repeat sequences was determined. We carried out phylogenetic analysis based on homologous protein-coding genes among 21 species derived from Fabaceae. We found that the phylogeny was largely congruent with prior hypotheses about the position of E. phaseoloides&nbsp;in evolutionary branches. The E. phaseoloides&nbsp;has a closer relationship with the Piptadeniastrum africanum.

Complete Chloroplast Genome and Comparative Analysis of Entada phaseoloides (Fabaceae)

<div>Background:&nbsp;The DNA barcoding is currently an effective and widely used tool that enables rapid and accurate identification of plant species.&nbsp; 
 
Methods:&nbsp;DNA barcoding of 9 chloroplast genes (rbcL, matK, trnH-psbA, accD-psaI, ndhA, psbE-petL, Rpl32-trnL, trnW-psaJ, trnSGCU-trnGGCC) were used to provide the theoretical basis for species identification, genetic diversity analysis of lotus population collected in Thua Thien Hue province, Vietnam. Universal primers were used and sequence products were analyzed using the MEGA X program. 
 
Result:&nbsp;The results showed that high levels of haplotype diversity (Hd), ranging from 0.618-0.869 and low levels of nucleotide diversity (Pi), ranging from 0.180 &times; 10-3-3.280 &times; 10-3 base on a total of nine gene regions of chloroplast genome. The neutrality tests show an excess of rare nucleotide position variations in individuals&rsquo; white lotus and derived haplotypes recent expansion. While the evolution of the individuals in the pink lotus may have to decrease. The phylogenetic analyses indicated that combined sequences were not insufficient to make a difference to the DNA barcoding in the individual&rsquo;s lotus of the N. nucifera species this is in the study. The standardized and accurate barcode information of lotus is provided for researchers. It lays the foundation for the conservation, evaluation, innovative utilization and protection of Nelumbonaceae germplasm resources.</div>

Research on Phylogenetic Relationship of Lotus Populations Collected in Thua Thien Hue Province, Vietnam based on the Chloroplast Genome by DNA Barcode