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Brittle stalk 2 encodes a putative glycosylphosphatidylinositol-anchored protein that affects mechanical strength of maize tissues by altering the composition and structure of secondary cell walls

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Abstract

A spontaneous maize mutant, brittle stalk-2 (bk2-ref), exhibits dramatically reduced tissue mechanical strength. Reduction in mechanical strength in the stalk tissue was highly correlated with a reduction in the amount of cellulose and an uneven deposition of secondary cell wall material in the subepidermal and perivascular sclerenchyma fibers. Cell wall accounted for two-thirds of the observed reduction in dry matter content per unit length of the mutant stalk in comparison to the wildtype stalk. Although the cell wall composition was significantly altered in the mutant in comparison to the wildtype stalks, no compensation by lignin and cell wall matrix for reduced cellulose amount was observed. We demonstrate that Bk2 encodes a Cobra-like protein that is homologous to the rice Bc1 protein. In the bk2-ref gene, a 1 kb transposon-like element is inserted in the beginning of the second exon, disrupting the open reading frame. The Bk2 gene was expressed in the stalk, husk, root, and leaf tissues, but not in the embryo, endosperm, pollen, silk, or other tissues with comparatively few or no secondary cell wall containing cells. The highest expression was in the isolated vascular bundles. In agreement with its role in secondary wall formation, the expression pattern of the Bk2 gene was very similar to that of the ZmCesA10, ZmCesA11, and ZmCesA12 genes, which are known to be involved in secondary wall formation. We have isolated an independent Mutator-tagged allele of bk2, referred to as bk2-Mu7, the phenotype of which is similar to that of the spontaneous mutant. Our results demonstrate that mutations in the Bk2 gene affect stalk strength in maize by interfering with the deposition of cellulose in the secondary cell wall in fiber cells.

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Abbreviations

Bk-2:

Brittle stalk-2

CesA:

Cellulose synthase

GPI:

Glycosylphosphatidylinositol

MPSS:

Massively parallel signature sequencing

Mu:

Mutator

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Authors and Affiliations

  1. DuPont Crop Genetics Research, DuPont Experimental Station Building E353, Route 141 and Henry Clay Road, Wilmington, DE, 19880-0353, USA

    Ada Ching, Timothy M. Bourett, Richard J. Howard & Antoni Rafalski

  2. Crop Genetics Research and Development, Pioneer Hi-Bred International, Inc., 7300 NW 62nd Avenue, P.O. Box 1004, Johnston, IA, 50131-1004, USA

    Kanwarpal S. Dhugga, Laura Appenzeller & Robert Meeley

Authors
  1. Ada Ching
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  2. Kanwarpal S. Dhugga
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  3. Laura Appenzeller
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  4. Robert Meeley
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  5. Timothy M. Bourett
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  6. Richard J. Howard
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  7. Antoni Rafalski
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Correspondence to Antoni Rafalski.

Appendix

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Table 3

Table 3 GenBank sequence submissions
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Ching, A., Dhugga, K.S., Appenzeller, L. et al. Brittle stalk 2 encodes a putative glycosylphosphatidylinositol-anchored protein that affects mechanical strength of maize tissues by altering the composition and structure of secondary cell walls. Planta 224, 1174–1184 (2006). https://doi.org/10.1007/s00425-006-0299-8

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