Comprehensive Transcriptome Profiling Uncovers Molecular Mechanisms and Potential Candidate Genes Associated with Heat Stress Response in Chickpea
Abstract
:1. Introduction
2. Results
2.1. Generation, Mapping, and Assessment of RNA-Seq Reads
2.2. Distribution of DEGs across Six Chickpea Genotypes Contrasting for Heat Stress Response
2.3. Genes Consistently Displaying Significant Differential Expression across all Six Chickpea Genotypes
2.4. Genes That Show Significant Regulation in Tolerant Genotypes but Are Not Substantially Regulated in the Sensitive Genotypes
2.5. Functional Categorization of DEGs
2.6. Differentially Expressed Transcription Factor (TF) Families
2.7. Candidate Genes Underlying QTLs Governing Heat Tolerance in Chickpea
3. Discussion
4. Materials and Methods
4.1. Plant Material, Growth Conditions, and Heat Stress Treatment Imposition
4.2. RNA Extraction, Illumina Sequencing, and Quality Check of the Sequenced Reads
4.3. Alignment of RNA-Seq Reads to the Chickpea Reference Genome
4.4. Identification of DEGs, GO Enrichment and Pathway Analyses
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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S. No. | Sample ID | Accession | Stage | Tissue | Treat-ment | Total Reads | Filtered Reads | Filtered Reads (%) | Mapped Reads | Mapped Reads (%) |
---|---|---|---|---|---|---|---|---|---|---|
1 | 92944_BFL_C | ICCV 92944 | Vegetative | Leaf | Control | 10,535,724 | 10,396,767 | 98.68 | 10,200,059 | 98.11 |
2 | 92944_BFL_S | Vegetative | Leaf | Stress | 9,748,624 | 9,586,491 | 98.34 | 9,420,776 | 98.27 | |
3 | 92944_BFR_C | Vegetative | Root | Control | 9,219,200 | 9,109,880 | 98.81 | 8,903,056 | 97.73 | |
4 | 92944_BFR_S | Vegetative | Root | Stress | 7,212,904 | 7,145,108 | 99.06 | 6,941,474 | 97.15 | |
5 | 92944_AFL_C | Reproductive | Leaf | Control | 12,186,436 | 12,019,055 | 98.63 | 11,785,230 | 98.05 | |
6 | 92944_AFL_S | Reproductive | Leaf | Stress | 10,763,618 | 10,651,458 | 98.96 | 10,462,620 | 98.23 | |
7 | 92944_AFR_C | Reproductive | Root | Control | 8250,522 | 8,127,931 | 98.51 | 7,808,259 | 96.07 | |
8 | 92944_AFR_S | Reproductive | Root | Stress | 5,524,034 | 5,385,805 | 97.50 | 5,014,912 | 93.11 | |
9 | 1356_BFL_C | ICC 1356 | Vegetative | Leaf | Control | 13,092,636 | 12,902,779 | 98.55 | 12,623,893 | 97.84 |
10 | 1356_BFL_S | Vegetative | Leaf | Stress | 7,848,062 | 7,762,543 | 98.91 | 7,631,234 | 98.31 | |
11 | 1356_BFR_C | Vegetative | Root | Control | 8,840,168 | 8,731,055 | 98.77 | 8,530,613 | 97.70 | |
12 | 1356_BFR_S | Vegetative | Root | Stress | 7,331,290 | 7,249,290 | 98.88 | 7,071,163 | 97.54 | |
13 | 1356_AFL_C | Reproductive | Leaf | Control | 13,595,606 | 13,428,551 | 98.77 | 13,186,265 | 98.20 | |
14 | 1356_AFL_S | Reproductive | Leaf | Stress | 14,108,618 | 13,902,500 | 98.54 | 13,657,584 | 98.24 | |
15 | 1356_AFR_C | Reproductive | Root | Control | 9,227,816 | 9,070,316 | 98.29 | 8,845,298 | 97.52 | |
16 | 1356_AFR_S | Reproductive | Root | Stress | 11,544,052 | 11,227,552 | 97.26 | 11,026,008 | 98.20 | |
17 | 15614_BFL_C | ICC 15614 | Vegetative | Leaf | Control | 10,665,666 | 10,509,803 | 98.54 | 10,299,447 | 98.00 |
18 | 15614_BFL_S | Vegetative | Leaf | Stress | 9,627,154 | 9,487,221 | 98.55 | 9,310,571 | 98.14 | |
19 | 15614_BFR_C | Vegetative | Root | Control | 9,227,608 | 8,942,970 | 96.92 | 8,672,250 | 96.97 | |
20 | 15614_BFR_S | Vegetative | Root | Stress | 9,642,178 | 9,537,244 | 98.91 | 9,229,981 | 96.78 | |
21 | 15614_AFL_C | Reproductive | Leaf | Control | 8,068,362 | 7,934,679 | 98.34 | 7,776,680 | 98.01 | |
22 | 15614_AFL_S | Reproductive | Leaf | Stress | 7,523,960 | 7,432,553 | 98.79 | 7,246,601 | 97.50 | |
23 | 15614_AFR_C | Reproductive | Root | Control | 4,552,154 | 4,432,821 | 97.38 | 4,332,522 | 97.74 | |
24 | 15614_AFR_S | Reproductive | Root | Stress | 8,145,728 | 8,044,853 | 98.76 | 7,394,860 | 91.92 | |
25 | 5912_BFL_C | ICC 5912 | Vegetative | Leaf | Control | 7,969,184 | 7,869,032 | 98.74 | 7,618,997 | 96.82 |
26 | 5912_BFL_S | Vegetative | Leaf | Stress | 7,941,190 | 7,856,097 | 98.93 | 7,716,168 | 98.22 | |
27 | 5912_BFR_C | Vegetative | Root | Control | 10,438,074 | 10,085,541 | 96.62 | 9,785,839 | 97.03 | |
28 | 5912_BFR_S | Vegetative | Root | Stress | 8,414,738 | 8,336,666 | 99.07 | 8,141,952 | 97.66 | |
29 | 5912_AFL_C | Reproductive | Leaf | Control | 10,500,550 | 10,363,949 | 98.70 | 10,182,627 | 98.25 | |
30 | 5912_AFL_S | Reproductive | Leaf | Stress | 5,180,758 | 5,117,767 | 98.78 | 4,961,472 | 96.95 | |
31 | 5912_AFR_C | Reproductive | Root | Control | 6,801,290 | 6,728,029 | 98.92 | 6,557,444 | 97.46 | |
32 | 5912_AFR_S | Reproductive | Root | Stress | 11,591,808 | 11,410,935 | 98.44 | 11,130,213 | 97.54 | |
33 | 4567_BFL_C | ICC4567 | Vegetative | Leaf | Control | 12,433,794 | 12,216,153 | 98.25 | 11,994,301 | 98.18 |
34 | 4567_BFL_S | Vegetative | Leaf | Stress | 10,425,446 | 10,288,128 | 98.68 | 10,068,543 | 97.87 | |
35 | 4567_BFR_C | Vegetative | Root | Control | 9,393,346 | 9,167,255 | 97.59 | 8,885,865 | 96.93 | |
36 | 4567_BFR_S | Vegetative | Root | Stress | 5,773,594 | 5,725,855 | 99.17 | 5,541,685 | 96.78 | |
37 | 4567_AFL_C | Reproductive | Leaf | Control | 12,591,206 | 12,398,886 | 98.47 | 12,166,369 | 98.12 | |
38 | 4567_AFL_S | Reproductive | Leaf | Stress | 9,909,176 | 9,778,957 | 98.69 | 9,596,564 | 98.13 | |
39 | 4567_AFR_C | Reproductive | Root | Control | 13,244,230 | 13,065,481 | 98.65 | 12,701,915 | 97.22 | |
40 | 4567_AFR_S | Reproductive | Root | Stress | 7,681,168 | 7,497,257 | 97.61 | 7,175,938 | 95.71 | |
41 | 10685_BFL_C | ICC 10685 | Vegetative | Leaf | Control | 10,826,998 | 10,674,415 | 98.59 | 10,455,869 | 97.95 |
42 | 10685_BFL_S | Vegetative | Leaf | Stress | 11,806,122 | 11,612,804 | 98.36 | 11,332,255 | 97.58 | |
43 | 10685_BFR_C | Vegetative | Root | Control | 11,425,388 | 11,082,467 | 97.00 | 10,756,797 | 97.06 | |
44 | 10685_BFR_S | Vegetative | Root | Stress | 7,308,052 | 7,241,501 | 99.09 | 7,058,819 | 97.48 | |
45 | 10685_AFL_C | Reproductive | Leaf | Control | 12,762,970 | 12,546,086 | 98.30 | 12,296,546 | 98.01 | |
46 | 10685_AFL_S | Reproductive | Leaf | Stress | 10,978,522 | 10,853,672 | 98.86 | 10,661,986 | 98.23 | |
47 | 10685_AFR_C | Reproductive | Root | Control | 7,721,326 | 7,641,066 | 98.96 | 7,464,657 | 97.69 | |
48 | 10685_AFR_S | Reproductive | Root | Stress | 9,251,526 | 9,124,015 | 98.62 | 8,884,704 | 97.38 | |
Total | 458,852,576 | 451,701,239 | 98.45 | 440,508,881 | 97.41 |
Gene ID | XLOC ID | Log2 Fold Change (Treatment/Control) | Gene Description | |||||
---|---|---|---|---|---|---|---|---|
ICCV 92944 | ICC 1356 | ICC 15614 | ICC 5912 | ICC 4567 | ICC 10685 | |||
Leaf sample—before flowering | ||||||||
Ca_14776 | XLOC_001145 | −2.161 | −2.165 | −2.125 | −1.702 | −1.651 | 1.532 | pathogenesis-related protein 10 |
Ca_07845 | XLOC_009218 | −2.956 | 2.458 | −2.134 | −0.806 | 1.402 | −0.163 | adenylate isopentenyltransferase |
Leaf sample—after flowering | ||||||||
Ca_00611 | XLOC_000503 | 3.140 | −3.321 | 2.981 | −0.057 | 0.576 | 1.887 | basic 7S globulin |
Ca_00716, Ca_00717 | XLOC_007162 | −2.600 | 2.662 | 2.696 | 0.615 | 1.725 | −0.349 | IST1-like protein isoform X2 |
Ca_05434 | XLOC_010012 | −2.650 | 2.103 | 4.128 | −0.786 | 1.774 | −0.593 | protein trichome birefringence-like 36 |
Ca_10969 | XLOC_012559 | −2.618 | 2.066 | 2.539 | 1.526 | 1.894 | 0.050 | selenoprotein H |
Ca_11344 | XLOC_014356 | −2.875 | 2.951 | 2.332 | 0.728 | 1.912 | NA | nuclear transcription factor Y subunit C-4 |
Ca_25834 | XLOC_017871 | 3.984 | 2.594 | 2.935 | 0.960 | 0.517 | 1.430 | glycine-rich cell wall structural 1-like |
Ca_05233 | XLOC_019232 | −2.424 | 2.341 | 2.077 | 1.273 | 0.213 | −0.392 | beta-xylosidase/alpha-L-arabinofuranosidase 2 |
Ca_06616 | XLOC_022893 | −2.089 | 2.078 | 2.323 | 0.228 | 1.379 | −0.698 | DUF936 family protein |
Ca_01970 | XLOC_025377 | −2.259 | 2.122 | 2.621 | 1.089 | 1.683 | 0.103 | chalcone-flavanone isomerase family |
Ca_27831 | XLOC_030374 | −2.519 | 2.197 | 2.352 | 0.991 | 1.566 | 1.025 | CTP synthase-like isoform X1 |
Ca_00464 | XLOC_000429 | 2.041 | −3.878 | −3.219 | 0.674 | NA | −0.680 | serine carboxypeptidase-like 11 |
Ca_06899 | XLOC_002705 | 2.490 | −2.172 | −2.465 | −0.001 | −0.193 | 1.441 | NAC family transcription factor 4 |
Ca_09600 | XLOC_018907 | 2.256 | −2.235 | −2.951 | −0.953 | −0.489 | −0.075 | transcription factor TGA4-like isoform X1 |
Ca_02987 | XLOC_021039 | 2.420 | −9.719 | −4.680 | 0.607 | −0.407 | 1.176 | ABA-responsive protein ABR18-like |
Root sample—before flowering | ||||||||
Ca_14147 | XLOC_000968 | 4.027 | 2.467 | 2.073 | 1.012 | NA | 1.305 | cysteine-rich repeat secretory protein 38-like |
Ca_03546 | XLOC_009482 | 3.704 | 3.129 | 2.302 | 1.785 | 1.538 | 0.734 | peroxidase 5-like |
Ca_06976, Ca_06977 | XLOC_001062 | −2.850 | −2.721 | −2.895 | 0.538 | 0.289 | −0.728 | UDP-glycosyltransferase 79B30 |
Ca_06902 | XLOC_002703 | −3.674 | 2.910 | −2.009 | 0.407 | −1.651 | 1.136 | PREDICTED: uncharacterized protein LOC101506036 |
Ca_14738 | XLOC_002759 | −2.465 | −2.426 | −2.305 | −1.309 | −1.193 | NA | E3 ubiquitin- ligase LIN-1 |
Ca_19160 | XLOC_003847 | 2.254 | −2.392 | −3.952 | −1.107 | −0.690 | −1.771 | arabinogalactan peptide 21-like |
Ca_18792 | XLOC_006264 | −2.067 | −2.650 | −3.088 | −0.820 | −1.456 | −0.846 | dirigent protein 9-like |
Ca_16612 | XLOC_010291 | −2.866 | −2.265 | −3.618 | −1.585 | −1.241 | −0.737 | probable pectate lyase 1 |
Ca_23747 | XLOC_013214 | −2.464 | −2.961 | −2.093 | −1.327 | NA | 0.074 | probable pectinesterase/pectinesterase inhibitor 25 |
Ca_22983 | XLOC_026437 | −2.194 | 3.352 | −2.720 | 1.481 | −1.969 | 1.948 | hypothetical protein glysoja_025159 |
Root sample—after flowering | ||||||||
Ca_11978 | XLOC_007090 | −2.016 | −2.800 | 2.140 | −0.071 | −0.706 | −0.682 | ras-related protein RABE1c-like |
Ca_18112 | XLOC_003876 | 2.567 | 2.437 | −2.526 | 1.302 | −1.692 | 1.597 | mitochondrial uncoupling protein 5 |
Ca_09783 | XLOC_005932 | 2.229 | −2.034 | −2.205 | −0.125 | NA | −0.189 | OBERON-like protein |
Ca_05504 | XLOC_009968 | −2.783 | −2.225 | −2.072 | −0.018 | −0.801 | −0.283 | glutamine--tRNA ligase |
Ca_24258 | XLOC_022284 | −3.033 | 2.178 | −3.424 | 0.380 | NA | 0.127 | NA |
NA, not available |
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Kudapa, H.; Barmukh, R.; Garg, V.; Chitikineni, A.; Samineni, S.; Agarwal, G.; Varshney, R.K. Comprehensive Transcriptome Profiling Uncovers Molecular Mechanisms and Potential Candidate Genes Associated with Heat Stress Response in Chickpea. Int. J. Mol. Sci. 2023, 24, 1369. https://doi.org/10.3390/ijms24021369
Kudapa H, Barmukh R, Garg V, Chitikineni A, Samineni S, Agarwal G, Varshney RK. Comprehensive Transcriptome Profiling Uncovers Molecular Mechanisms and Potential Candidate Genes Associated with Heat Stress Response in Chickpea. International Journal of Molecular Sciences. 2023; 24(2):1369. https://doi.org/10.3390/ijms24021369
Chicago/Turabian StyleKudapa, Himabindu, Rutwik Barmukh, Vanika Garg, Annapurna Chitikineni, Srinivasan Samineni, Gaurav Agarwal, and Rajeev K. Varshney. 2023. "Comprehensive Transcriptome Profiling Uncovers Molecular Mechanisms and Potential Candidate Genes Associated with Heat Stress Response in Chickpea" International Journal of Molecular Sciences 24, no. 2: 1369. https://doi.org/10.3390/ijms24021369