<div>Background: Bituminaria bituminosa is a long-day plant. The plant has a high pastoral value and palatability. It is grazed by cattle, goats and sheep. Leaves and stems contain four furanocoumarins&rsquo; psoralen, angelicin, xanthotoxin and bergapten. The seed is covered by the fruit. Germination was not observed in beaked fruits. A cut with a scalpel can increase the germination rate. According to the literature there are not enough studies about the culture media and convenient conditions. 
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Methods: In our study, we tried six medium for callus production with different hormone concentrations including 2,4-D, BAP and kinetin. In this study, a protocol for callus induction and analysis of isoflavonoids and terpenes of Bituminaria bituminosa has been developed. 
 
Result: Maximum callus induction frequency was observed on Murashige and Skoog (MS) medium supplemented with 0.75 mg/L 2,4-D + 2.5 mg/L kinetin in young primary leaves. 0.75 mg/L 2,4-D+2.5 mg/L kinetin medium is more successful in the production ofses of flavonoids that can be produced in vitro and is more successful in producing phenols. In terpene production, it is seen that 1.25 mg/L 2,4-D+3.5 mg/L BAP, which is the best callus producing medium containing BAP, is the most successful medium among all tested mediums.</div>

Callus Production and Analysis of Isoflavonoids and Some Terpenes from Bituminaria bituminosa

<div style="text-align: justify;">Legumes, in general, are rich in glycosidic isoflavone conjugates, whereby specific b-glucosidases act to release the free aglycones that could serve in plant-microbe interactions and defense. Prosopis cineraria is an important legume tree, bearing medicinally useful and edible pods, generally growing in extreme arid/semi-arid zones of Rajasthan. However, specific phytochemical-cum-metabolic profiles are lacking for the same. Therefore, the present investigation undertook phytochemical and metabolic screening of the pods/seeds of P. cineraria for the presence of putative isoflavonoids viz. &nbsp;genistein and daidzein, their glycosides and b-glucosidase(s) capable of catalyzing the glycoside hydrolysis. &nbsp;Extraction and identification of &nbsp;these &nbsp;two &nbsp;aglycone forms of &nbsp;the above &nbsp;isoflavonoids &nbsp;were &nbsp;performed &nbsp;with &nbsp;solvent &nbsp;partition chromatography &nbsp;and &nbsp;Fluorescent/High &nbsp;Performance Thin Layer Chromatography, respectively. Furthermore, optimization of the isoflavone conjugate-specific b-glucosidase activity with respect to pH optima, temperature and time were carried out. The partially purified enzyme showed a temperature optima of 50&deg;C and pH optima of 4.5. The enzyme also demonstrated activity towards natural substrates daidzin and genistin which are glycosides of isoflavonoids daidzein and genistein respectively. The methanolic extracts of the seeds of P. cineraria indicated the presence of related isoflavonoids which needs to be further validated.</div>

Identification of an isoflavone glycoside hydrolyzing b-glucosidase in the seeds of the desert legume Prosopis cineraria

Background: Harvesting period is an important feature that not only affects the yield quality of Astragalus herbs but also the content of astragaloside IV and calycosin 7-O-&beta;-D-glucopyranoside which&nbsp; are considered as the core indicators of the quality of Astragalus herbs. Herb yield and quality undergo rapid and substantial changes during the harvesting process. Part of enzymes and their encoding genes have been identified as potential candidates for the biosynthesis of herb quality-related triterpenoid saponins and isoflavonoids. However, wide gap still exists in the understanding of the pathways involving growth, development and biosynthesis of these two important active components in the roots of Astragalus. 
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Methods: In this study, we used Astragalus roots as material to analyze its metabolome and transcriptome of herbs at harvesting stage. 
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Result: The results indicated that changes in the harvesting period had a significant effect on the yield and quality of the herbs. Correlation of metabolome and RNA-seq results constructed using weighted gene co-expression network analysis (WGCNA) with herb growth and development indicated that the expression patterns of 19 hub metabolites (Oxamniquine, 4-ene-Valproic acid, Traumatic Acid, L-Histidinol, Salidroside, Temsavir, Retaspimycin, Isosakuranetin, etc.) and 16 structural genes with annotations (CNX2, talA, ABCF2, HIBCH, crtZ, etc.) had the highest connectivity with root growth and secondary metabolism of Astragalus. To search for potential regulatory genes related to root growth and development, we screened 54 relevant genes using 16 hub genes with annotations. The results advance the molecular basis of biosynthesis of secondary metabolites related to the harvesting period of Astragalus herbs, which can provide a theoretical basis for determining the regulatory mechanism of yield and quality.

Weighted Correlation Network Analysis Mining Key Metabolites and Genes for Regulation of Root Growth and Secondary Metabolism of Astragalus membranaceus mongholicus