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      저자 : Himanshu Tak (검색결과 2 건)
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        Overexpression of MusaNAC68 reduces secondary wall thickness of xylem tissue in banana

        Sanjana Negi,Himanshu Tak,T. R. Ganapathi 한국식물생명공학회 2019 Plant biotechnology reports Vol.13 No.2

        Secondary wall is an important component of xylem tissue, as it maintains the integrity of the vessel elements and provides mechanical strength for upward growth of plants. Enough prior evidence has linked secondary wall thickening with auxinsignaling pathway. Hence, plants showing auxin-mediated effects might demonstrate alteration of secondary wall depositions. GUS expression from promoter of NAC68 (pMusaNAC68) was observed in many organs with strong expression in vascular tissues. Transgenic banana plants overexpressing NAC68 transcription factor were analyzed for lignin and secondary wall depositions. Strong reduction of lignin in cross-sections of transgenic lines was observed by phloroglucinol-HCl and toluidine blue-O staining. The reduction of secondary wall thickness observed after calcofluor white staining was substantiated by data from scanning electron microscopy indicating reproducibly lower secondary wall depositions. A strong reduction in transcript levels of PAL, 4CL, C4H, COMT and CcOAMT was observed due to the overexpression of NAC68. Furthermore, substantial elevation of MYB transcription factors such as MYB4a-like and MYB4b-like which are probable repressors of phenylpropanoid biosynthesis pathway was also observed. Expression of MYB transcription factors, MYB85-like and MYB58-like which are probable activators of secondary wall depositions, was remarkably downregulated after NAC68 overexpression. Transgenic lines also showed altered expression of genes coding for cellulose synthase subunits with remarkable elevation in expression of a close homologue of Arabidopsis CesA8, which functions in cellulose deposition during secondary wall development. The study indicated a novel function of NAC68 transcription factor in regulating secondary wall thickening and will enhance our knowledge about crosstalk between auxin-signaling and secondary wall depositions.

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        In vitro xylem vessel elements formation from banana embryogenic cells and expression analysis of vessel development-related genes

        Sanjana Negi,Himanshu Tak,T. R. Ganapathi 한국식물생명공학회 2015 Plant biotechnology reports Vol.9 No.2

        Tracheary elements formation was observed in banana embryogenic cells cultured in brassinolide-supplemented medium after a period of 10 days. The highest frequency of tracheary elements observed after 10 days was up to 31.88 %. The auxin, 2,4-D was found to be inhibitor of tracheary elements formation in brassinolidesupplemented medium and at a concentration of 1.5 lM of 2,4-D, the frequency of tracheary elements formation was reduced to zero. The appearance of secondary wall in these xylem vessel elements was observed by toluidine blue staining and lignin autofluorescence under fluorescence as well as laser confocal microscopy. Similar to lignin autofluorescence imaging, the calcofluor white staining also revealed development of characteristic secondary wall in these xylem vessel elements indicating deposition of cellulose along with lignin during secondary wall formation. The xylem vessel elements induced comprised of fiber-like elements (FLEs) and tracheary elements with deposition of secondary wall. Expression analysis of lignin biosynthesis pathway genes indicated that important genes like CCR, CCoAOMT and COMT were induced as early as fifth day of brassinolide (BL) treatment. Changes in transcript level of key master regulators of xylogenesis during tracheary elements formation showed that expression of vascularrelated NAC domain containing transcription factors (VND) were up-regulated with induction of VND1–3 and VND6–7 expression as early as third day of brassinolide treatment. This study of in vitro tracheary elements formation in embryogenic cell suspension cultures is useful in understanding the genes involved in secondary wall thickening in banana.

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