Rather than by direct acquisition via lateral gene transfer, GHF5 cellulases were passed on from early Pratylenchidae to root-knot and cyst nematodes

Table 1 Overview of PCR primers used for cellulase amplication from individual nematodes

A
CD1	CD2
TATPPPYGQLSVSGTKLVDSSGQPVQLIGNSLFWHQFQAQYWNAETVKALKCNWNANVVRAAVGVDLERGYMSDP
ENG1
TTAYNQAVAVIEAAISQGLYVIVDWHSHESHVDKAIEFFTKIAKAYGSYPHVLYETFNEPLQGVSWTDILVPYHKKVIAAI
CD4	ENG2
RALDSKNVIILGTPTWCQDVDIASQNPIKEYKNLMYTFHFYAATHFVNGLGAKLQTAINNGLPIFVTEYGTCSADGNGNI
CD6
DTNSISSWWSLMDNLKISYLNWAISDKSETCSALKPGTPAANVGVSSSWTTSGNMVADHDKKKSTGVSCS
B Primer sequence 5’→3’ *		Primer sequence 5’→3’ *
Region PPYGQLS (CD1):		Region WCQDV (CD4):
CD1aF	ccIccItacggIcaattgtc	CD4aR	tccacRtcctgggacca
CD1bF	ccIccItatggIcaattgtc	CD4cR	tccacAtcttggcacca
CD1cF	ccIccItatggIcaattatc
CD1PraFa	ccgccgtatgggcaa	Region FVTEYG (ENG2):
CD1PraFb	cctccctatggccaa	ENG2 see e.g.[22]	gtIccRtaYTcIgtIacRaa
CD1PraFc	cgccgtatgggcaa
CD1MelF	ctccatatgggcaattatctgt	Region ISYLNWAISD (CD6):
Region LKCNWN (CD2):		CD6PraFb	tctcctacatcaactgggc
CD2aF	ctcaaatgcaattggaacKc	CD6aR	gcccagttggcgtaIgaga
CD2bF	ctcaaatgcaattggaatKc	CD6bR	gcccaIttggcRtaIgaaa
CD2cF	cttaaatgcaIttggaatKc	CD6cR	gcccaIttgaIgtaMgaaa
CD2dF	cttaaatgctIttggaatKc	CD6dR	gcccagttgaYgtaIgaga
Region YVIVDW (ENG1):		CDGp8R	gcccagttgaggtacgaa
ENG1 see e.g.[22]	taYgtIatcgtIgaYtggca	CD6PraRa	cccagttggcgtagga
ENG1R	tgccaRtcIacgatIacRta	CD6MelR	tgtttgagatagcccagttg

* I=inosine; K= g or t, M= a or c, R= a or g, Y= c or t. In bold: discriminative nucleotide position.
Conserved amino acid motives in GHF5 cellulases from plant parasitic nematodes residing in nematode Clade 12 (Holterman et al. 2006 [24]). Primer design was based on these motives, and all primers used in this study are listed bellow. A. The backbone sequence given below is derived from the predicted amino acid sequence of the potato cyst nematode (Globodera rostochiensis) cellulase Gr-eng-1 (GenBank AF004523), amino acid positions 18 – 324 (mainly catalytic domain). Underlined: part of signal peptide for secretion. B. Primer names and primer sequences.

ISSN: 2730-7182