Table 1.
List of primers used in the study.
Fig 1.
Schematic illustration of the T-DNA vector used for maize transformation.
Pyruvate orthophosphate dikinase (PDK) intron of 767bp introduced between the sense and antisense chitinase sequence was driven by poly Ubi maize promoter and TMV omega enhancer to increase transgene expression.
Fig 2.
Phylogenetic tree analysis showing the relationship of C. partellus chitinase genes among lepidopteran chitinase.
Analysis was based on the neighbour-joining method according to amino acid sequences using MEGA X. Bootstrap support values with 1,000 samples are shown on the branches.
Fig 3.
Kaplan-Meier analysis indicating survival after feeding purified dsCHI and bacterially-induced dsCHI to C. partellus larvae.
Fig 4.
The effect of dsCHI on the various phenotype (A) various deformed shapes of dsRNA fed larvae compared to those feed without dsRNA (control), (B) different morphological deformity observed in some pupa from dsRNA fed larvae compared to the normal pupa (control), C) phenotype abnormality resulted from dsRNA fed larva compared to larvae that was fed with control.
Fig 5.
Relative knockdown in transcript levels of chitinase gene during in-vitro feeding assay with bacterially expressed dsCHI after (A) period of 5 days post-exposure. (B) period of 15 days post-exposure. Values are expression mean ±standard error and different letters indicate significant different (p<0.05) between dsCHI gene and the controls (Empty HT115 and dsGFP).
Fig 6.
Maize genomic DNA and amplification of dsCHI.
(A) λ indicate lambda ladder and 1–8 lanes indicated quality of genomic DNA isolated from Maize. (B) L is 1kb plus ladder and 1 lane does not show amplification from non-transgenic Maize while 2–8 lanes showed the PCR amplification of genomic DNA from 7 events.
Fig 7.
Histochemical staining of transgenic and control maize.
(A) Blue colour indicating the presence of GUS gene in germinated embryo. (B) Blue-like colour indicated the presence of GUS gene in spikelets.
Fig 8.
RT-qPCR analysis of T1 transgenic maize plants.
Muta1—Muta6 indicating different transgenic lines. Values are expression mean ± standard error and different letter indicate significant different (p < 0.05).
Fig 9.
(A) Indicating the area non-transgenic maize consume larva (B) indicated the area of transgenic maize consume by larva (C) illustrating the quantification of the area consumed per larva at 4 days of feeding.
Fig 10.
(A) Relative growth rate comparison estimates between larvae that were fed with non-transgenic Maize (wc) and transgenic maize leaves(w+) (B) dsCHI transcript abundance in C. partellus larvae after dsCHI-transgenic and non-transgenic post feeding. values are mean ±standard error and different letters indicate significant different (p<0.05).