. 2020 Aug 13;10:13713. doi: 10.1038/s41598-020-70530-z

Table 3.

Growth and culm quality characteristics of bovine lysozyme expressing sugarcane lines.

Bovine lysozyme (BvLz_m) expressing line	Bvlz_m yield (mg/kg culm fresh mass)	Agronomic parameter
Bovine lysozyme (BvLz_m) expressing line	Bvlz_m yield (mg/kg culm fresh mass)	Leaf length (cm)	Culm height (cm)	Tiller number	Culm biomass (fresh mass, kg/plant)	Brix (%)
Non-transformed (CP72-1210)	0.0 ± 0.0^a	75.3 ± 1.4^a	12.5 ± 0.2^a	4.0 ± 0.2^a	7.1 ± 0.3^a	14.6 ± 0.6^a
*Single promoter maizeubiquitin 1:BvLz***_m expressing lines (CP72-1210)
33	0.22 ± 0.01^b*	74.0 ± 0.5^a	13.5 ± 0.2^a	10.0 ± 0.4^b**	9.1 ± 0.6^b*	17.2 ± 0.6^b*
67	0.27 ± 0.01^b*	72.3 ± 1.1^a	14.5 ± 0.6^a	4.0 ± 0.3^a	7.8 ± 0.5^a	15.7 ± 0.5^b*
108	0.33 ± 0.01^b*	76.8 ± 1.4^a	18.0 ± 0.3^b**	5.0 ± 0.2^a	9.6 ± 0.9^b*	14.9 ± 0.4^a
114	0.32 ± 0.01^b*	70.5 ± 2.2^a	11.3 ± 0.6^a	6.0 ± 0.2^b*	8.2 ± 0.6^b*	16.8 ± 0.4^b*
116	0.36 ± 0.01^b*	74.0 ± 3.3^a	14.5 ± 1.0^a	8.0 ± 0.9^b**	7.0 ± 0.3^a	17.9 ± 0.7^b*
123	0.26 ± 0.01^b*	76.5 ± 1.8^a	17.1 ± 1.3^b*	4.0 ± 0.1^a	8.9 ± 0.5^b*	15.2 ± 0.5^a
*Triple promoter:BvLz***_m expressing lines
pUPE:BvLz_m 32 (CP72-1210)	2.7 ± 0.1^b**	81.9 ± 1.2^a	16.3 ± 0.4^b*	17.0 ± 2.9^b**	13.1 ± 1.2^b*	18.4 ± 0.3^b*
pUDE:BvLz_m (TCP98-4454)
18	5.1 ± 0.3^b**	92.3 ± 4.1^b**	17.3 ± 0.2^b*	7.0 ± 0.6^b*	10.5 ± 0.9^b*	16.7 ± 0.3^b*
19	4.6 ± 0.2^b**	95.6 ± 2.5^b**	18.5 ± 0.4^b**	7.0 ± 0.2^b*	11.3 ± 1.0^b*	15.6 ± 0.4^a
44	2.9 ± 0.1^b**	74.8 ± 2.6^a	14.3 ± 1.4^a	7.0 ± 1.3^b*	8.6 ± 0.8^b*	16.0 ± 0.2^b*
54	6.0 ± 0.3^b**	87.2 ± 4.0^b*	16.6 ± 1.1^b*	4.0 ± 0.5^a	10.7 ± 1.1^b*	17.9 ± 0.6^b*
*Quadruple promoter:BvLz***_m expressing lines
pUPBE:BvLz_m (TCP98-4454)
1	6.7 ± 0.3^b**	115.3 ± 3.1^b**	19.4 ± 1.2^b**	21.1 ± 1.3^b**	16.4 ± 0.7^b**	19.1 ± 0.6^b*
2	10.0 ± 0.7^b**	201.8 ± 6.4^b**	32.1 ± 2.4^b**	17.0 ± 1.1^b**	20.8 ± 1.0^b**	18.0 ± 0.7^b*
4	8.3 ± 0.4^b**	140.2 ± 5.1^b**	25.3 ± 1.9^b**	14.0 ± 0.9^b**	18.6 ± 0.9^b**	17.5 ± 0.3^b*

Morphological parameters (leaf height, culm height and tiller number) of 15 representative single promoter:bovine lysozyme (BvLz_m) and five representative triple promoter:BvLz_m expressing lines were measured every 2 weeks for 4 months after planting. Culm biomass was determined at 11 months (harvest). The BvLz_m yield is indicated as determined by enzyme-linked immunosorbent assay in juice extract of culms at the 11-month harvest (one kg of culm). Total soluble solids (Brix) of extracted culm juice from 11-month-old plants was determined using a refractometer (model PR-101α, Atago U.S.A, Inc., Bellevue, WA). Data represent means from four biological replications ± standard errors. Means are compared column-wise. Values that are significantly different from those of non-transformed at p < 0.001 and p < 0.0001 are denoted by * and **, respectively. BvLz_m: maize codon-optimized BvLz; pUPE:BvLz_m: BvLz_m expressed from three promoters, maize ubiquitin 1 (pUbi), sugarcane proline-rich protein (pSHPRP) and sugarcane elongation factor 1α (pSHEF1α); pUDE:BvLz_m: BvLz_m expressed from three promoters, pUbi, sugarcane dirigent16 and pSHEF1α. pUPBE:BvLz_m: BvLz_m expressed from four promoters, pUbi, pSHPRP, pSHEF1α and Sugarcane bacilliform virus promoter.