增强UV—B辐射对高山植物麻花艽净光合速率的影响

在高寒矮嵩草（Kobresia humilis)草甸地区以太阳短波辐射为背景,建立了人工增强UV－B辐射的实验装置,每天增补15．8kJ.m^-2的辐射剂量,模拟平流层臭氧破坏约5％时近地表面太阳UV－B辐射的增强。观测表明：UV－B辐射的增强对麻花艽（Gentiana stramianea)植物的光合作用无明显的抑制或伤害作用。相反,在早晨补充UV－B辐射的短时间内,叶片的Pn随Gs的增大而有所提高。随着UV－B辐射时间的延长,在11：30－12：30,Pn和Gs有所降低。UV－B辐射时间进一步延长后（约14：00以后）,处理和对照组叶片Pn和Gs的差异趋向不明显,增强太阳UV－B辐射后,麻花艽叶片的光合色素并无明显变化,UV－B吸收物质的含量无明显变化,麻花艽叶片厚度的直接测量表明,增强UV－B辐射能明显提高叶片的厚度。叶片厚度的增加可补偿增强UV－B辐射辐后引起的光合色素的光降解,改善单位叶面积为基础的光合速率,是高原植物对强UV－B辐射的一种适应方式。     

重金属镉（Cd）和增强UV—B辐射复合对大豆生长和生理代谢的影响

研究了大豆的生长、生物量、抗氧化酶活性和吲哚乙酸（IAA）氧化酶活性在Cd^2 、UV－B辐射和二者复合胁迫（Cd UV-B)下的变化。结果表明,Cd^2 和UV－B辐射都抑制大豆生长,并显著抑制根的伸长,二者复合后加强了对根伸长的抑制。UV－B辐射显著增强了POD、SOD活性,Cd^2 对POD活性影响不明显,但却拮抗UV－B对POD活性的诱导,SOD活性在各种胁迫下显著增强。虽然Cd%2 对叶片类黄酮含量影响不明显,但对UV－B诱导的类黄酮合成有一定影响。IAA氧化酶活性在复合作用下下降,可能是复合胁迫影响大豆生长的重要因素之一。     

环丁烷嘧啶二聚体累积与水稻UV—B敏感性的关系

利用单克隆抗体ELISA ,研究了UV_B对水稻DNA中环丁烷嘧啶二聚体 (CPD)的诱导形成及其光、暗修复 ,并对CPD累积与水稻UV_B敏感性的关系进行了分析。结果表明 ,我国南方的 5个水稻 (OryzasativaL .)品种经13.6kJ·m-2 ·d-1UV_B处理 15d后 ,在株高、生物量、光合作用等方面表现出明显的品种间差异。不同品种水稻的DNA中CPD累积比对照明显增加 ,且敏感品种CPD的累积比抗性品种显著提高。统计分析证实 ,CPD的累积与生物量的抑制呈显著正相关 (r2 =0 .6 2 2 )。UV_B诱导的水稻DNA中CPD的清除以光修复为主 ,不同品种CPD暗修复能力相似 ,而光修复能力存在明显差异。根据以上结果推测 ,不同水稻品种UV_B敏感性与CPD光修复能力的差异有关。     

UV-B辐射对香蕉光合作用和不同氮源利用的影响

生长在NO3^--N、NH4^--N和NH4NO3－N的香蕉叶片有相近似的最大光合速率,UV－B辐射引起生长在不同氮源的香蕉叶片光合速率、表现量子产率和光肥利用效率的降低。UV－B辐射使生长在不同氮源的植株叶面积干重和叶氮含是降低。生长在NH4^--N的植株Vcmax和Jmax均较生长在其它氮源的高。UV－B辐射引起生长在NH4^-N的植株Vcmax和Jmax降低较相同处理的NO3^-－N和NH4NO3－N植株明显,表明生长在NH4^ －N的香蕉对UV－B辐射更加敏感。UV－B辐射改变植株的叶片的碳氢比和碳氮比。经过UV－B辐射处理的NH4^ －N生长植株的碳氮生长在NO3^--N和NH4NO3－N的低。UV－B辐射可能改变植株对不同氮源的吸收利用,从而引起碳氮代谢和酸碱调节的变化。UV－B辐射降低叶氮在Rubisco和生物力能学组分的分配系数,可能使这些组分合成减少,使叶片光调节的变化。UV－B辐射降低叶氮在Rubisco和生物力能学组分的分配系数,可能使这些组分合成减少,使叶片光合速率下降。结果表明,生长在不同氮源的香蕉植树对UV－B辐射有不同响应,NH4^ -N有利于主要光合参数增高,但其对UV－B辐射亦最为敏感。氮供应受限制或植株生长在中性盐如NH4NO3－N则对UV－B辐射不甚敏感。     

芒果老叶在增强UV-B辐射处理下的损伤和保护反应

以‘台衣一号’芒果盆栽苗离体老叶为试材,研究增强UV—B辐射条件下芒果老叶的损伤和保护反应。结果表明：UV—B辐射处理使芒果叶片MDA含量和相对电导率升高、叶绿素含量和叶绿素a／b降低,表明叶片受到损伤,且随处理时间延长叶片损伤加重。UV—B辐射处理叶片可溶性蛋白含量、抗氧化酶（SOD、CAT、POD）活性、保护色素（类胡萝卜素、类黄酮）和还原型GSH含量显著高于对照叶片,UV—B辐射处理叶片维生素C含量显著低于对照叶片,表明增强UV—B辐射可诱导叶片细胞通过提高活性氧清除能力和积累保护色素而直接吸收部分UV—B辐射来提高抗增强UV—B辐射损伤的能力。     

温度对UV-B诱导的烟草叶圆片DNA损伤的影响

环丁烷嘧啶二聚体(CPD)和6-4光产物(6-4PP)是两种主要的UV-B诱导的DNA光损伤产物。利用单克隆抗体酶联免疫吸附分析法(ELISA),研究了温度对UV-B诱导的烟草叶圆片DNA损伤的影响。室温(24℃)条件下,UV-B处理引起了烟草叶圆片DNA中CPD和6-4PP的积累。0℃条件下,UV-B处理的烟草叶圆片DNA中CPD和6-4PP的积累比室温下分别降低了9.8%和12%。UV-B诱导的DNA损伤曾被认为是纯粹的光化学过程而与不受温度影响,而本实验结果表明,UV-B诱导的烟草叶圆片DNA形成CPD和6-4PP的过程具有温度依赖性。这一特性有利于植物对全球变化的适应,因而具有重要的生态学意义。     

大豆作物响应增强UV—B辐射的品种差异

田间条件下模拟20％平流层臭氧层衰减,紫外线（UV－B,280－315nm)辐射增强,研究了UV－B对2个大豆（Glycin max (L.)Merr.)品种黑豆和晋豆生长,光合作用和稳定碳同位素组成的影响,结果表明,晋豆比黑豆对UV－B有较强的抗性或不敏感,表现为增强的UV－B辐射显著抑制黑豆的生长和株高,叶、茎、根和总生物量以及株高全部降低,而晋豆仅茎重和株高降低;晋豆的色素含量（叶绿素a,b,类胡萝卜素和类黄酮）不受UV－B辐射影响,在UV－B辐射下黑豆的净光合作用,气孔导度,胞间CO2浓度和蒸腾作用以及不分利用效率明显下降,而晋豆只有气孔导度和蒸腾作用减少,这可能与晋豆本身含有较高的类黄酮及较多的表皮毛和遗传特性有关,用叶片稳定碳同位素组成（δ^13C值）的分析也证明晋豆对UV－B辐射不敏感,由此看来,大豆品种对UV－B辐射的反应差异可以通过δ^13C值来判定。     

不同强度的UV-B辐射对高山植物麻花艽光合作用及暗呼吸的影响

以人工种植的多年生高山植物麻花艽(Uentiana straminea)为材料,在3个不同强度的UV—B辐射处理下,定时测定处理和对照叶片的净光合速率、表观量子效率和暗呼吸的变化。结果显示：UV—B处理对麻花艽叶片的光合作用在短期内有一定的抑制作用,但随着处理时间的增加,该高山植物能很快地适应强UV—B辐射的处理。表明麻花艽这种青藏高原常见的高山植物在长期的自然选择过程中可能已经形成了适应UV—B辐射的特有生理机制。暗呼吸的实验结果亦表明：在3种强度的UV—B辐射处理下,麻花艽叶片的呼吸作用从一开始就未受到抑制;随着UV—B辐射时间的增加,UV—B辐射强度越高,呼吸强度越强;这可能是UV—B辐射并未引起麻花艽呼吸机构的破坏所致。     

UV—A区段紫外线照射对DNA影响的拉曼光谱分析

本文检测了鲱鱼精DNA水溶液经不同时间UV－A紫外辐射后的拉曼光谱,研究结果表明,该区段紫外辐射比用UV－A和UV－B共同照射对DNA的影响要小,主链构象基本稳定。但经较长时间辐射仍会对鲱鱼精DNA造成损伤,受影响的部位主要是脱氧核糖和胸腺嘧啶碱基部分,UV－A对脱氧核糖的影响与UV－A加UV－B共同照射的结果作比较后,可以说明UV－A对脱氧核糖的损伤有累积的效应,而对于胸腺嘧啶的影响,从其各个指标的分析来看,有损伤但程度较小。本实验说明UV－A辐射条件下没有嘧啶二聚体的形成,也不存在6,4光产物形成的证明,但对于Dewar异构体的形成,有部分证明,与Taylor(1994)报道的结果相一致,UV－A没有造成DNA单链断裂现象。     

增强UV-B辐射与干旱复合处理对小麦幼苗生理特性的影响

为研究由于平流层臭氧层减薄紫外线B辐射增强在干旱地区对春小麦生理特性的影响的特殊性,模拟平流层臭氧减少20％时辐射到地表的紫外线B（UV－B,280－315nm)的增强和水分胁迫（－0.5Mpa,聚乙二醇PEG－6000处理获得）,通过测定两种胁迫下春小麦（Triticum aestivum L.)叶绿素含量、类黄酮含量、水势、细胞膜相对透性、超氧歧化酶（SOD）活性及丙二醛（MDA）含量,研究了增强UV－B辐射和水分胁迫复合作用对温室种植的小麦幼苗生理生化的影响。实验结果表明,虽然水分胁迫和UV－B辐射单独或复合处理都使春小麦的叶绿素含量降低,但仅UV－B辐射增强单独处理显著地降低小麦叶绿素a、b和总叶绿素的含量,而水分胁迫以及复合处理对叶绿素的含量的降低作用不显著。两种胁迫无论是单独作用还是复合作用均能使类黄酮含量升高,并且处理第3天比第1天高出近50％,复合处理下类黄酮的含量大于两个因子单独处理。UV－B辐射和水分胁迫处理1d对春小麦叶片的相对电导率的影响不明显,处理3d后两种胁迫下相对电导率均上升,表明膜透性增加,其中水分胁迫作用下增加尤其明显。膜质过氧化产物丙二醛的含量,在两种因子单独和复合作用下都升高,说明膜的生理功能受到了一定的不利影响。活性氧清除剂超氧化物歧化酶（SOD）的活性在各种处理下,都没有发生改变。虽然增强的UV－B辐射和干旱处理一样,会显著降低植物水势,但是,UV－B辐射与干旱同时处理时叶片水势降低的程度,不但没有比两者分别处理时降低程度之和低,而且比单做干旱处理时的降低程度还低,这表明UV－B辐射和干旱胁迫同时处理时,UV－B辐射不但没有加重水分胁迫,反而减轻了干旱对春小麦生长的胁迫。由此可以认为,在干旱条件下,增强UV－B辐射不会加剧而是有利于提高小麦对干旱的抗性。     

The native cyclobutane pyrimidine dimer photolyase of rice is phosphorylated

The cyclobutane pyrimidine dimer (CPD) is a major type of DNA damage induced by ultraviolet B (UVB) radiation. CPD photolyase, which absorbs blue/UVA light as an energy source to monomerize dimers, is a crucial factor for determining the sensitivity of rice (Oryza sativa) to UVB radiation. Here, we purified native class II CPD photolyase from rice leaves. As the final purification step, CPD photolyase was bound to CPD-containing DNA conjugated to magnetic beads and then released by blue-light irradiation. The final purified fraction contained 54- and 56-kD proteins, whereas rice CPD photolyase expressed from Escherichia coli was a single 55-kD protein. Western-blot analysis using anti-rice CPD photolyase antiserum suggested that both the 54- and 56-kD proteins were the CPD photolyase. Treatment with protein phosphatase revealed that the 56-kD native rice CPD photolyase was phosphorylated, whereas the E. coli-expressed rice CPD photolyase was not. The purified native rice CPD photolyase also had significantly higher CPD photorepair activity than the E. coli-expressed CPD photolyase. According to the absorption, emission, and excitation spectra, the purified native rice CPD photolyase possesses both a pterin-like chromophore and an FAD chromophore. The binding activity of the native rice CPD photolyase to thymine dimers was higher than that of the E. coli-expressed CPD photolyase. These results suggest that the structure of the native rice CPD photolyase differs significantly from that of the E. coli-expressed rice CPD photolyase, and the structural modification of the native CPD photolyase leads to higher activity in rice.     

Increase in CPD photolyase activity functions effectively to prevent growth inhibition caused by UVB radiation

Rice cultivars vary widely in their sensitivity to ultraviolet B (UVB) and this has been correlated with cyclobutane pyrimidine dimer (CPD) photolyase mutations that alter the structure/function of this photorepair enzyme. Here, we tested whether CPD photolyase function determines the UVB sensitivity of rice (Oryza sativa) by generating transgenic rice plants bearing the CPD photolyase gene of the UV-resistant rice cultivar Sasanishiki in the sense orientation (S-B and S-C lines) or the antisense orientation (AS-D line). The S-B and S-C plants had 5.1- and 45.7-fold higher CPD photolyase activities than the wild-type, respectively, were significantly more resistant to UVB-induced growth damage, and maintained significantly lower CPD levels in their leaves during growth under elevated UVB radiation. Conversely, the AS-D plant had little photolyase activity, was severely damaged by elevated UVB radiation, and maintained higher CPD levels in its leaves during growth under UVB radiation. Notably, the S-C plant was not more resistant to UVB-induced growth inhibition than the S-B plant, even though it had much higher CPD photolyase activity. These results strongly indicate that UVB-induced CPDs are one of principal causes of UVB-induced growth inhibition in rice plants grown under supplementary UVB radiation, and that increasing CPD photolyase activity can significantly alleviate UVB-caused growth inhibition in rice. However, further protection from UVB-induced damage may require the genetic enhancement of other systems as well.     

Higher amounts of anthocyanins and UV-absorbing compounds effectively lowered CPD photorepair in purple rice (Oryza sativa L.)

Growth of a near‐isogenic line (NIL) for the purple leaf gene Pl of rice with a genetic background of Taichung 65 (T‐65) rice was significantly retarded by supplementary ultraviolet‐B radiation (UV‐B), despite the fact that the amounts of UV‐absorbing compounds and anthocyanins in NIL were significantly higher than those in T‐65. In order to understand the role of flavonoids in UV‐B induced damage protection in T‐65 and the NIL, both the (1) relationships between changes in the steady state of cyclobutane pyrimidine dimer (CPD) levels and changes in accumulation of anthocyanins and UV‐absorbing compounds in leaves with leaf age, and (2) the susceptibility to CPD induction by UV‐B radiation and the ability to photorepair CPD were examined. Although supplementary UV‐B elevated the steady state of CPD levels in leaves in both strains, the level in the leaf of the NIL was higher than that in T‐65 at any time. The susceptibility to CPD induction by short‐term (challenge) UV‐B exposure was lower in the NIL than in T‐65. On the other hand, the CPD photorepair was also lower in the leaves of the NIL than in those of T‐65. The decrease in CPD‐photorepair in the NIL was due to a lowering of the leaf‐penetrating blue/UV‐A radiation, which is effective for photoreactivation by photolyase, by anthocyanins. Thus, accumulation of anthocyanins and UV‐absorbing compounds did not effectively function as screening against damage caused by elevated UV‐B radiation in the NIL, and the retardation of growth in the NIL resulted from its lower ability to photorepair CPD by higher amounts of anthocyanins.     

DNA repair in higher plants; photoreactivation is the major DNA repair pathway in non-proliferating cells while excision repair (nucleotide excision repair and base excision repair) is active in proliferating cells

We investigated expression patterns of DNA repair genes such as the CPD photolyase, UV-DDB1, CSB, PCNA, RPA32 and FEN-1 genes by northern hybridization analysis and in situ hybridization using a higher plant, rice (Oryza sativa L. cv. Nipponbare). We found that all the genes tested were expressed in tissues rich in proliferating cells, but only CPD photolyase was expressed in non-proliferating tissue such as the mature leaves and elongation zone of root. The removal of DNA damage, cyclobutane pyrimidine dimers and (6–4) photoproducts, in both mature leaves and the root apical meristem (RAM) was observed after UV irradiation under light. In the dark, DNA damage in mature leaves was not repaired efficiently, but that in the RAM was removed rapidly. Using a rice 22K custom oligo DNA microarray, we compared global gene expression patterns in the shoot apical meristem (SAM) and mature leaves. Most of the excision repair genes were more strongly expressed in SAM. These results suggested that photoreactivation is the major DNA repair pathway for the major UV-induced damage in non-proliferating cells, while both photoreactivation and excision repair are active in proliferating cells.     

UVB sensitivity and cyclobutane pyrimidine dimer (CPD) photolyase genotypes in cultivated and wild rice species.

We investigated the UVB-sensitivity in 12 rice strains belonging to two cultivated species (O. sativa and O. glaberrima) and three wild species (O. barthii, O. meridionalis and O. rufipogon) of rice possessing the AA genome, while focusing on the CPD photolyase activity and the genotypes of CPD photolyase. Although the UVB sensitivity, CPD photolyase activity, and CPD photolyase genotype varied widely among these rice species, the sensitivity to UVB radiation depended on the activity of the CPD photolyase, regardless of grass shape, habitat, or species. The rice strains examined here clearly divided into three groups based on the CPD photolyase activity, and the activity of the strains greatly depended on amino acid residues at positions 126 and 296, with the exception of the W1299 strain (O. meridionalis). The amino acid residues 126 and 296 of CPD photolyase in Sasanishiki strain (O. sativa), which showed higher enzymatic activity and more resistance to UVB, were glutamine (Gln) and Gln, respectively. An amino acid change at position 126 from Gln to arginine ("Nori"-type) in the photolyase led to a reduction of enzymatic activity. Additionally, an amino acid change at position 296 from Gln to histidine led to a further reduction in activity. The activity of the W1299 strain, which possesses a "Nori"-type CPD photolyase, was the highest among the strains examined here, and was similar to that of the Sasanishiki. The CPD photolyase of the W1299 contains ten amino acid substitutions, compared to Sasanishiki. The alterations in amino acid residues in the W1299 CPD photolyase compensated for the reduction in activity caused by the amino acid substitutions at positions 126. Knowledge of the activity of different CPD photolyase genotypes will be useful in developing improved rice cultivars.     

Spontaneously occurring mutations in the cyclobutane pyrimidine dimer photolyase gene cause different sensitivities to ultraviolet-B in rice

Sensitivity to ultraviolet-B (UVB) radiation (280-320 nm) varies widely among rice cultivars. We previously indicated that UV-resistant rice cultivars are better able to repair cyclobutane pyrimidine dimers (CPDs) through photorepair than are UV-sensitive cultivars. In this paper, we report that UVB sensitivity in rice, in part, is the result of defective CPD photolyase alleles. Surjamkhi (indica) exhibited greater sensitivity to UVB radiation and was more deficient in CPD photorepair ability compared with UV-resistant Sasanishiki (japonica). The deficiency in CPD photorepair in Surjamkhi resulted from changes in two nucleotides at positions 377 and 888 in the photolyase gene, causing alterations of two deduced amino acids at positions 126 and 296 in the photolyase enzyme. A linkage analysis in populations derived from Surjamkhi and Sasanishiki showed that UVB sensitivity is a quantitative inherited trait and that the CPD photolyase locus is tightly linked with a quantitative trait locus that explains a major portion of the genetic variation for this trait. These results suggest that spontaneously occurring mutations in the CPD photolyase gene cause different degrees of sensitivity to UVB in rice, and that the resistance of rice to UVB radiation could be increased by increasing the photolyase function through conventional breeding or bioengineering.     

Ultraviolet-B sensitivities in Japanese lowland rice cultivars: cyclobutane pyrimidine dimer photolyase activity and gene mutation

There is a cultivar difference in the response to ultraviolet-B(UVB: 280–320 nm) in rice (Oryza sativa L.). AmongJapanese lowland rice cultivars, Sasanishiki, a leading Japaneserice cultivar, is resistant to the damaging effects of UVB whileNorin 1, a close relative, is less resistant. We found previouslythat Norin 1 was deficient in cyclobutane pyrimidine dimer (CPD)photorepair ability and suggested that the UVB sensitivity inrice depends largely on CPD photorepair ability. In order toverify that suggestion, we examined the correlation betweenUVB sensitivity and CPD photolyase activity in 17 rice cultivarsof progenitors and relatives in breeding of UV-resistant Sasanishikiand UV-sensitive Norin 1. The amino acid at position 126 ofthe deduced amino acid sequence of CPD photolyase in cultivarsincluding such as Norin 1 was found to be arginine, the CPDphotolyase activities of which were lower. The amino acid atthat position in cultivars including such as Sasanishiki wasglutamine. Furthermore, cultivars more resistant to UVB werefound to exhibit higher photolyase activities than less resistantcultivars. These results emphasize that single amino acid alterationfrom glutamine to arginine leads to a deficit of CPD photolyaseactivity and that CPD photolyase activity is one of the mainfactors determining UVB sensitivity in rice. ¹ These authors contributed equally to the paper. ² Corresponding author: E-mail, kumagai{at}ige.tohoku.ac.jp; Fax,+81-22-217-5691.     

Cyclobutane pyrimidine dimer (CPD) photolyase repairs ultraviolet-B-induced CPDs in rice chloroplast and mitochondrial DNA

Plants use sunlight as energy for photosynthesis; however, plant DNA is exposed to the harmful effects of ultraviolet‐B (UV‐B) radiation (280–320 nm) in the process. UV‐B radiation damages nuclear, chloroplast and mitochondrial DNA by the formation of cyclobutane pyrimidine dimers (CPDs), which are the primary UV‐B‐induced DNA lesions, and are a principal cause of UV‐B‐induced growth inhibition in plants. Repair of CPDs is therefore essential for plant survival while exposed to UV‐B‐containing sunlight. Nuclear repair of the UV‐B‐induced CPDs involves the photoreversal of CPDs, photoreactivation, which is mediated by CPD photolyase that monomerizes the CPDs in DNA by using the energy of near‐UV and visible light (300–500 nm). To date, the CPD repair processes in plant chloroplasts and mitochondria remain poorly understood. Here, we report the photoreactivation of CPDs in chloroplast and mitochondrial DNA in rice. Biochemical and subcellular localization analyses using rice strains with different levels of CPD photolyase activity and transgenic rice strains showed that full‐length CPD photolyase is encoded by a single gene, not a splice variant, and is expressed and targeted not only to nuclei but also to chloroplasts and mitochondria. The results indicate that rice may have evolved a CPD photolyase that functions in chloroplasts, mitochondria and nuclei, and that contains DNA to protect cells from the harmful effects of UV‐B radiation.     

Eukaryotic class II cyclobutane pyrimidine dimer photolyase structure reveals basis for improved ultraviolet tolerance in plants

Ozone depletion increases terrestrial solar ultraviolet B (UV-B; 280–315 nm) radiation, intensifying the risks plants face from DNA damage, especially covalent cyclobutane pyrimidine dimers (CPD). Without efficient repair, UV-B destroys genetic integrity, but plant breeding creates rice cultivars with more robust photolyase (PHR) DNA repair activity as an environmental adaptation. So improved strains of Oryza sativa (rice), the staple food for Asia, have expanded rice cultivation worldwide. Efficient light-driven PHR enzymes restore normal pyrimidines to UV-damaged DNA by using blue light via flavin adenine dinucleotide to break pyrimidine dimers. Eukaryotes duplicated the photolyase gene, producing PHRs that gained functions and adopted activities that are distinct from those of prokaryotic PHRs yet are incompletely understood. Many multicellular organisms have two types of PHR: (6-4) PHR, which structurally resembles bacterial CPD PHRs but recognizes different substrates, and Class II CPD PHR, which is remarkably dissimilar in sequence from bacterial PHRs despite their common substrate. To understand the enigmatic DNA repair mechanisms of PHRs in eukaryotic cells, we determined the first crystal structure of a eukaryotic Class II CPD PHR from the rice cultivar Sasanishiki. Our 1.7 Å resolution PHR structure reveals structure-activity relationships in Class II PHRs and tuning for enhanced UV tolerance in plants. Structural comparisons with prokaryotic Class I CPD PHRs identified differences in the binding site for UV-damaged DNA substrate. Convergent evolution of both flavin hydrogen bonding and a Trp electron transfer pathway establish these as critical functional features for PHRs. These results provide a paradigm for light-dependent DNA repair in higher organisms.