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Although the chlorophyll (Chl) degradation gene\n                    <jats:italic>LpNOL<\/jats:italic>\n                    negatively regulates both processes, its upstream regulatory mechanism remains elusive. In this study, a bHLH transcription factor LpCbDR2 was identified as a direct activator of\n                    <jats:italic>LpNOL<\/jats:italic>\n                    through yeast one\u2010hybrid screening. LpCbDR2 binds the G\u2010box\n                    <jats:italic>cis<\/jats:italic>\n                    \u2010element within the proximal\n                    <jats:italic>LpNOL<\/jats:italic>\n                    promoter, as demonstrated by EMSA,\n                    <jats:italic>in planta<\/jats:italic>\n                    transactivation, and CUT&amp;Tag\u2010qPCR assays. Expression analysis reveals that\n                    <jats:italic>LpCbDR2<\/jats:italic>\n                    transcript levels positively correlate with\n                    <jats:italic>LpNOL<\/jats:italic>\n                    expression during leaf development and in response to heat stress. Functional characterization using overexpression and RNAi transgenic lines demonstrates that LpCbDR2 negatively regulates both heat tolerance and leaf senescence. Overexpression lines show severer degradation of photosystem complex proteins, increased reactive oxygen species (ROS) accumulation, reduced antioxidant enzyme activities, and lower survival rates under heat stress, whereas RNAi lines exhibit opposite phenotypes. Epistatic analysis by suppressing both\n                    <jats:italic>LpCbDR2<\/jats:italic>\n                    and\n                    <jats:italic>LpNOL<\/jats:italic>\n                    confirmed that LpCbDR2 acts upstream of\n                    <jats:italic>LpNOL<\/jats:italic>\n                    . Importantly, RNAi\u2010\n                    <jats:italic>LpCbDR2<\/jats:italic>\n                    lines accumulate 35.2\u201354.4% higher crude protein contents without yield penalty. These findings establish LpCbDR2 as a key regulator of\n                    <jats:italic>LpNOL<\/jats:italic>\n                    that links heat tolerance and senescence pathways, making it a potential target for enhancing stress tolerance and feedstock quality in perennial ryegrass and other forage species.\n                  <\/jats:p>","DOI":"10.1111\/tpj.71007","type":"journal-article","created":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T16:33:06Z","timestamp":1782405186000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A\n                    <scp>bHLH<\/scp>\n                    transcription factor,\n                    <scp>LpCbDR2<\/scp>\n                    , activates\n                    <i>LpNOL<\/i>\n                    to regulate senescence, heat tolerance, and crude protein content in perennial ryegrass"],"prefix":"10.1111","volume":"126","author":[{"given":"Tingchao","family":"Yin","sequence":"first","affiliation":[{"name":"Sanya Institute of Nanjing Agricultural University  Sanya 572024 People's Republic of China"},{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyi","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huanhuan","family":"Hao","sequence":"additional","affiliation":[{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuwei","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Zhou","sequence":"additional","affiliation":[{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3779-1603","authenticated-orcid":false,"given":"Aying","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Life Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3547-3476","authenticated-orcid":false,"given":"Yingjun","family":"Chi","sequence":"additional","affiliation":[{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9701-7583","authenticated-orcid":false,"given":"Bin","family":"Xu","sequence":"additional","affiliation":[{"name":"Sanya Institute of Nanjing Agricultural University  Sanya 572024 People's Republic of China"},{"name":"College of Agro\u2010grassland Science Nanjing Agricultural University  Nanjing 210095 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