Supplementary Material: File 1

Figure 1

Representative gel pictures showing differential expression pattern between Control/mock treated (C) and P.personata treated (T) samples. Differential expression pattern was observed using Genefishing DEG kit, involving isolation of total RNA from control (C) and pathogen treated (T) leaves, cDNA synthesis followed by amplification of cDNA with different random arbitrary primers (See Material and Methods for details). Arrows indicate the differentially upregulated bands in pathogen treated (T) samples compared with control(C).

Figure 2A

A pie diagram showing the primary role known for/assigned to the proteins encoded by the genes isolated in this study.

Figure 2B

A pie diagram illustrating the number of isolated clones with reported role in plant stress or defense responses.

Figure 3

Pair wise sequence alignment of DIR1 and DIR2 nucleotide sequences.

Table 1

Sequence information of gene specific primers utilized in this study.

Name / Forward / Reverse
PAL / GGAACCGGTATGCTCACC / GCATCATGTATTTCATTGGCA
C4H / GCGAAATCAACGAAGACAAC / CCAACGGCAATGACTTCAG
CAD / GTTCACTACTGTGGACTCC / CGGGTTTATGCATAGCACC
DIR1 / CTACACAATGGTGACACAAC / AGTCAAATGCAGTTGTATTAAC
DIR2 / CTACACAATGGTGACACAAC / GCAGTTGTATTTACTTTATACG
ACTIN / TGGCATCACACTTTCTACAA / CAACGGAATCTCTCAGCTCC

Established role of some of the cloned genes in plant defense.

Functional homologs of some of the cloned genes in other plant systems with established role in defense includeADP-ribosylation factor (ARF)(AdDR-1), which was shown to be up-regulated in responded to fungal and elicitor treatment. Constitutively overexpressed rice ARF1 triggered spontaneous lesion mimic phenotype, induction of pathogenosis-related (PR) genes, reduced susceptibility to a fungal pathogen, and caused accumulation of SA (Lee et al. 2003). ADP-ribosylation factor induces cell death upon overexpression in N. benthamiana and found to be a component of non-host resistance and R-gene mediated resistance (Coemans et al. 2008). Sec61α subunit(AdDR-4), which is part of Sec61 translocon complex, involved in the translocation of proteins to cross the endoplasmic reticulum (ER) membrane. It was shown that protein secretory pathway is required for systemic acquired resistance (SAR) and NPR1 regulates the expression of several secretory pathway genes including Sec61α subunit gene in Arabidopsis (Wang et al. 2005).T-DNA insertions in SEC61 αcompromised the plant's ability to efficiently secrete PR1 as well as impaired resistance against the pathogen Pseudomonas syringae (Wang et al. 2005).CBL-interacting protein kinase (CIPK)(AdDR-5), which along with calcineurin B-like protein (CBL) is emerging as a major calcium signaling network with only reported role in abiotic stress responses, it would be interesting to study its role in biotic stress responses. Papain-like cysteine protease(AdDR34) is a member of the cysteine proteases, whihc have emerged as key enzymes in the regulationof animal PCD. Rcr3(papain-likecysteine endoprotease) is specificallyrequired for the function of Cf-2, a Lycopersicon pimpinellifoliumgene bred into cultivated tomato (Lycopersicon esculentum) forresistance to Cladosporium fulvum(Krüger et al. 2002).Zinc finger proteins (AdDR-7) were shown to have role in defense against pathogens (Oh et al. 2005). Putative leucine rich repeat proteins (AdDR-10) are plant disease resistance genes (R-genes), which mediate resistance to pathogens harboring corresponding avirulence (Avr) gene (Dangl and Jones 2001).

Tonoplast intrinsic protein (TIP) (AdDR-12) was found to have role in H2O2 transport as well as detoxification of light induced ROS and suggested to have involved in intracellular signal propagation (Maurel et al. 2009). Enzyme 1-deoxy-D-xylulose 5-phosphate synthase (AdDR-17) is part of terpenoid biosynthesis pathway. Glutathione S-transferase(GST) (AdDR-19) was proposed to be involved in detoxification and controlling the progress of cell death during hypersensitive reaction (Wagner et al. 2002). Putative translation initiation protein (AdDR-20) was reported as akey element of the signal transduction pathway resulting in plant programmed cell death (Hopkins et al. 2008). Lysin motif-type receptor-like kinase (AdDR-31) plays a role in recognizing both friends and foes (Knogge and Scheel 2006). AdDR32 encodes Pyridoxine biosynthesis protein, Pyridoxine (Vitamin B6) is a strong antioxidant with potential role in the plant-pathogen defense response. Proline is the mostdiversely used osmolyte that accumulates in osmotically stressed organisms. Proline dehydrogenase (ProDH) (AdDR-33) was transiently induced both in tobacco and in Arabidopsis upon R. fascians infection(Simón-mateo et al. 2006). Heat shock protein 70 (HSP70) (AdDR-37) along with HSP90 were found to be essential components of hypersensitive response and non-host resistance in N. benthamiana plants (Kanzakiet al. 2003). Thaumatin like proteins (AdDR-11) are class-V pathogenesis related proteins with antimicrobial activity (van Loon et al. 2006). Lipid transfer proteins (LTPs) (AdDR-51) belong to PR-14 family and known for antifungal and anti microbial activity (van Loon et al. 2006).

References

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