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Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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Use the information provided in the experiment below to annotate/ label the figure. Background: Boll weevil is a serious pest of cotton crop. Effective control involves applications of chemical insecticides, increasing the cost of production and environmental pollution. The current genetically modified Bt crops have allowed great benefits to farmers but show activity limited to lepidopteran pests. This work reports on procedures adopted for integration and expression of a cry transgene conferring resistance to boll weevil and fall armyworm by using molecular tools. Four Brazilian cotton cultivars were microinjected with a minimal linear cassette generating 1248 putative lines. Complete gene integration was found in only one line (TO-34) containing one copy of crylla detected by Southern blot. Protein was expressed in high concentration at 45 days after emergence (dae), decreasing by approximately 50% at 90 dae. Toxicity of the cry protein was demonstrated in feeding bioassays revealing 56.7% mortality to boll weevil fed buds and 88.1% mortality to fall armyworm fed leaves. A binding of crylla antibody was found in the midgut of boll weevils fed on TO-34 buds in an immunodetection assay. The gene introduced into plants confers resistance to boll weevil and fall armyworm. Transmission of the transgene occurred normally to TI progeny. All plants showed phenotypically normal growth, with fertile flowers and abundant seeds. Results relating to figure: A total of 1248 putative transgenic seeds were collected from microinjected bolls of four Brazilian cultivars and sown in the greenhouse. At first, PTLs were analysed by PCR using three primer combinations. A few amplicons were verified in the cultivars and, even so, in one or two primer combinations, indicating incomplete integration of the gene. Only in cv. BRS 293 was at least one PTL (TO-34) identified in all primer combinations, indicating possible complete integration of crylla based on PCR reactions, mainly with the 5 F/2R primer combination, which generated an amplicon corresponding to the complete gene sequence (2.1 kb). A fragment of 0.44 kb (1 F/3R) collected from this line was purified, sequenced and analysed using BLAST tools. The results showed high homology between the sequences of crylla from BRS 293 T0-34 and the Bt cry1Ia gene deposited in the NCBI gene bank (DQ535488, E-value: 3e−63 to 1e−12), as can be seen in Fig. 3.

A
DQ535488 Bacillus thuringiensis Cry1la gene, complete cds S=274 E=6.8e-70
Color key for alignment scores
40-50
50-80
80-200
Query
Bauungana.e.com.co
10
131 140
|----
261
|---
20
150
270
<40
B
H01 ATTTTTTTCTCAACCTGTTGTTGGAGTATCAAGAGGTGGTTTCCATTGGCRATTTCCCACGCTCCCARTAGCTTCTGATAATTTTTACTATCCTGGTTATGCTGGAATTGGAACGCAGCTTCRAGTTTCA
H04
30
280
50
160
40
170
50
****************
100
180
300
Description
290
|‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒
60
310
190
70
H01 AGATCGTACAAATAAAGTTGAACATAGGATAGT-ACTATCAGCTGGA
H04 AGATCGTACARATACTATTGARCCTAATATAATTACTCARATCCCTCTTGTGRATGCTTTCA
150
200
80
ATCCAGAGGTGGTTTCCATTGGRAATTTCCCACGCTCCCARTAGCTTCTGATAATTTTTACTATCCTGGTTATGCTGGAATTGGRACACAGCTTCARGATTCA
90
210
200
220
100
230
>=200
Max Total Query
Accession
score score cover value
274 274 96% 7670 84 % 0053540
110
250
H01
GAGAATGAATTACCACCTGAAGCCACAGGACAGCCARATTATGRATCATATAGACATAGATTTTCCCATATAGGACTCATCTCAGCTTCCCATGTGRAGGCATTGGTTATTCTTGGACACATCGTAGTGC
H04 GAGARTGRATTACCACCTGARGCARCAGGACAACCARATTACGRATCATATAGACATAGACTTTCCCATATAGGACTCATCTCAGCTTCCCATGTGAAGGCATTGGTTATTCTTGGACACATCGTAGTGC
240
120
130
......
250
260
Figure 3. Comparison of the nucleotide sequences encoded by the crylla (DQ535488) gene with 0.44 bp cry1la-CNPA deposited in the NCBI gene
bank, by using BLAST analysis (A) and alignment of both sequences using the MultipleM sequence alignment program (B). H01 - crylla (DQ535488);
H04 - cry1la-CNPA. The asterisk (*) below each nucleotide indicates the conserved sequence.

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