陕西三种特种稻米氨基酸及品质分析

以陕西洋县产茉莉香米、红香米和绿香粳米等 3种特种稻米为材料 ,分析了氨基酸及其它品质指标。结果表明 ,茉莉香米、红香米和绿香粳米均含有 1 7种氨基酸 ,氨基酸总含量 (mg/ 1 0 0 g)分别为 730 0、72 70、74 90 ;除Trp外 ,7种必需氨基酸占氨基酸总量分别为 34 .6 %、34 .5 %、34 .8% ;Lys含量 (mg/ 1 0 0g)分别为 5 5 0、5 2 0、5 30 ;粗蛋白质含量分别是 9.94 %、9.2 3%、8.5 2 % ;Cu含量 (mg/ 1 0 0g)分别是 0 .6 2、0 .94、0 .79;Zn含量 (mg/ 1 0 0 g)分别是 1 .99、1 .75、2 .0 2 ;Fe含量 (mg/ 1 0 0 g)分别是 1 .5 5、3.70、4 .5 0 ;直链淀粉含量分别是 1 5 .6 %、3.7%、1 5 .1 % ;胶稠度分别是 33mm、4 0 .6mm、78mm ;糊化温度分别是 6 .1级、6 .8级、7级 ;水分含量分别是 1 1 .2 7%、1 1 .5 5 %、1 2 .5 2 %。这 3种特种稻品质较优 ,具有推广价值     

云南作物资源特征特性及生态地理分布研究Ⅰ.云南地方稻种资源中特种稻资源

对6121份云南地方稻种资源中的特种稻资源,即黑（紫）米、红米、香米、糯米（酒米）和软米的特征特性进行综合评价;以云南省128个县为基本单位,应用GIS平台统计整理云南省每个县所属的气候类型,并结合云南省的5个稻作分区分析研究云南特种稻资源的生态地理分布。通过研究综合评价了云南特种稻资源的特征特性,初步摸清了云南特种稻资源除香米资源集中分布在中亚热带和中温带外,其他特种稻资源都集中分布在南亚热带的生态地理分布习性,其研究结果可为稻作资源收集保护、农作物生态地理分布、特优稻育种、生态育种、生物育种、农业产业开发和产业区划结构调整等研究提供参考依据。     

几种同工酶与猕猴桃品种对溃疡病抗性关系的研究

应用聚丙烯酰胺凝胶电泳、同工酶分析技术分别研究了猕猴桃植株体内过氧化物酶(POD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、酯酶(EST)同工酶谱带的变化,结果表明:自然感染溃疡病前后,此6种同工酶谱带特征在不同抗感品种中表现出一定的差异.未感染溃疡病菌前,抗(感)品系枝条、叶片POD同工酶均有2条酶带,PPO同工酶有3条酶带,但感病品种酶带颜色深且粗,而抗病品种酶带颜色浅且细,叶片酶带颜色深于枝条;SOD、CAT同工酶谱带均为1条,Rf值分别为0.38、0.28,感性品种较抗耐品种谱带亮度高活性强;自然发病后,抗(感)品系POD、PPO同工酶谱带数都增加,分别为4、3条和5、4条,且抗病品种新酶带出现较感病品种早且酶带粗颜色深活性强,感病品系虽也有新酶带出现,但酶带少活性弱,抗病品系枝条、叶片POD、PPO同工酶新谱带的Rf值分别为0.63、0.67和0.85、0.87;抗感病品种SOD、CAT同工酶都被诱导产生了1条新的同工酶谱带,Rf分别为0.32和0.27,新酶带现色时间迟,且酶带颜色浅活性弱,但抗耐品种较感性品种谱带亮且活性强;EST同工酶于自然发病前后变化不大,与抗病性关系不很明显.     

紫色甘薯品系6种酶的同工酶分析

采用聚丙烯酰胺凝胶电泳技术,选用过氧化物酶(POD)、超氧化物歧化酶(SOD)、细胞色素氧化酶(CYT)、淀粉酶(AMY)、酯酶(EST)、苹果酸脱氢酶(MDH)等六种酶的同工酶系统对14个紫色甘薯品种(系)进行了同工酶分析.结果表明:过氧化物酶(POD)、细胞色素氧化酶(CYT) 、超氧化物歧化酶(SOD)等三种酶的同工酶酶谱很丰富,分别有13、11、10条酶带;淀粉酶(AMY)、酯酶(EST)、苹果酸脱氢酶(MDH)等三种酶的同工酶酶带相对较少,分别为4、7、7条.聚类分析结果表明,所有供试紫色甘薯品系或品种分成7类:I类包括品系A1、A2、A7;II类包括品系B1～B3;III类包括品系B7和B8;IV类只有品系B9;V类包括品系A3、A4、A6;VI类只有品系A5;VII类只有品种"山川紫".     

蚕豆种质资源清蛋白遗传多样性分析?

利用SDS-PAGE对101份蚕豆种质资源进行了清蛋白遗传多样性分析,共检测出蛋白带2625条。除共有带外,迁移率不同的谱带类型36种,其中相对分子量为92kD、75kD、62kD、40kD、34kD、17kD、13kD的谱带在检测的种质材料中出现的频率最高,分别为92.08%、90.10%、99.01%、95.05%、95.05%、98.02%、95.05%。其余29种谱带类型具有较强的多态性,多态性谱带平均为16.09条,多态性比例为44.69%。每份种质材料的清蛋白谱带数介于21～31条之间,平均25.99条;供试种质间遗传相似系数0.6111～0.9722,平均0.7122。3个地理类群内多样性指数0.9879,类群间多样性指数0.0121,表明蚕豆清蛋白的变异主要来源于类群内。聚类分析将参试种质分为6类,与以往蚕豆种质分类研究结果类似,表明清蛋白能在一定程度上反映种质间的亲缘关系。研究结果对于蚕豆蛋白质品质育种具有一定的参考价值。     

蝮蛇抗栓酶与精制蝮蛇抗栓酶制剂的比较研究

本文作者对以东北长白山白眉蝮蛇毒为原料生产的两种酶制剂——蝮蛇抗栓酶与精制蝮蛇抗栓酶进行了比较研究。用 HPLC 柱层析粗毒得到15个蛋白峰,同时对两种酶制剂以 HPLC 层析用两根层析柱串联上样层析以提高其分辩率,得到两个图谱,蝮蛇抗栓酶有7个蛋白峰,而精制品有3～4个蛋白峰.同时以 SDS-PAGE 电泳图谱.蝮蛇抗栓酶呈现7～8条谱带,精制品有4～5条谱带,并与已知分子量的标准蛋白进行对比,结果表明两种酶制剂均非单一组分.蝮蛇抗栓酶谱带较多,精制品较纯.其分子量均在2～6万之间.蝮蛇抗栓酶几个组分有协同作用,每次剂量在0.25～0.50单位,即有明显疗效,而精制品临床用药剂量较大,每次0.75～1.0单位,多者达1.25单位(3～5支).     

八倍体小冰麦及其亲本种子醇溶蛋白和高分子量麦谷蛋白亚基的研究

对５个八倍体小冰麦种子醇溶蛋白和高分子量麦谷蛋白亚基的电泳谱带进行了分析,结果表明：八倍体小冰麦中１和中２的电泳谱带基本相同,中３、中４、中５的电泳谱带基本相同,但完全不同于中１和中２的类型。八倍体小冰麦中１和中２同天蓝冰草（Ａｇｒｏｐｙｒｏｎｉｎｔｅｒｍｅｄｉｕｍ（Ｈｏｓｔ）Ｐ．Ｂ．＝Ｅｌｙｔｒｉｇｉａｉｎｔｅｒｍｅｄｉａ（Ｈｏｓｔ）Ｎｅｖｓｋｉ＝Ｔｈｉｎｏｐｙｒｕｍｉｎｔｅｒｍｅｄｉｕｍ（Ｈｏｓｔ）ＢａｒｋｗａｒｔｈａｎｄＤｅｗｅｙ）在高分子量麦谷蛋白亚基上存在一条相同的谱带,在醇溶蛋白谱带上出现了小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）和冰草均没有的带型。中３、中４、中５在醇溶蛋白谱带上具有一条冰草×染色体组的特征谱带,其基因表达程度同冰草类似。从５个八倍体小冰麦种子醇溶蛋白和高分子量麦谷蛋白亚基的电泳图谱结果,分析了八倍体小冰麦染色体组构成及亲本来源,并探讨了八倍体小冰麦在优质麦育种过程中的价值。     

草鱼呼肠孤病毒新分离株（GCRV991）的病毒学特性分析

从湖南长沙分离到一株致病性强的草鱼呼肠孤病毒 (GCRV991) ,该病毒能使草鱼CIK ,肥头鲤FHM细胞产生明显的CPE ,对水生动物BF2 ,EPC及哺乳动物BHK ,VERO细胞株不敏感。中和实验显示 ,GCRV873 抗体能有效地中和GCRV991病毒颗粒 ,形成抗原抗体免疫复合物。纯化的病毒核酸与蛋白经SDS PAGE分离 ,分别呈现 11条清晰的核酸带及 5条主要与 2条微量结构多肽图谱 ,其核酸蛋白分子量大小与GCRV873 相近似。该毒株基因组总分子量为 14.48× 10 6kD ,大小范围是 0 .5 5～ 2 .6 1× 10 6kD ;5条主要与两条微量结构多肽分子量近似值分别为136kD、132kD、6 5kD、43kD、34kD及 138kD、82kD。上述结果提示 ,新分离的GCRV991与GCRV873 具有相似的抗原性     

西藏豌豆资源蛋白质含量及遗传多样性分析

为探究西藏高原豌豆资源遗传多样性,分析种子蛋白质含量与生态地理环境间的关系。本研究搜集整理了54份来自西藏不同生态地理环境条件下的豌豆材料。利用SDS-PAGE对54份豌豆种子水溶性和盐溶性蛋白进行遗传多样性分析,测定种子蛋白质含量,分析地理生态因子(经度、纬度、海拔)与总蛋白含量间的关系。结果表明,54份豌豆材料种子总蛋白含量为17.58%~28.67%,其中水溶性蛋白占86.12%~91.40%,盐溶性蛋占4.76%~8.29%。相关分析表明,西藏豌豆种子总蛋白含量与经度呈显著正相关,与纬度呈正相关,而与海拔高度呈负相关。54份豌豆种子水溶性和盐溶性蛋白SDSPAGE分别检测出蛋白谱带1588条和699条,其中水溶性蛋白分出迁移率不同的谱带类型43种,基因多样性指数为0~0.50,相对分子量在24.71~149.54 Ku之间,且其谱带多集中于24.87~50.41 Ku低相对分子量区和56.34~88.08 Ku高相对分子量区;盐溶性蛋白分离出迁移率不同的谱带类型24条,基因多样性指数0~0.50,相对分子量在24.85~91.24 Ku之间。基于蛋白的谱带的聚类分析将54份豌豆资源可划分为7个类群,遗传相似系数变异范围为0.60~0.91;根据海拔高度将54份豌豆资源划分为4个地理类群,各类群蛋白谱带的基因多样性指数分别为0.23、0.18、0.35和0.31,Shannon's信息指数分别为0.33、0.41、0.52和0.46。西藏豌豆资源遗传变异较丰富,表现出随着海拔高度增加,遗传变异呈增大趋势。同时种子贮藏蛋白含量与经度呈显著正相关,而与纬度和海拔高度相关性不大。     

RAPD分子标记在酱油生产菌系统发育分析中的应用

以米曲霉沪酿3.042(AS3.951)、酱油曲霉AS3.495为参照,对从商品酱油曲精中分离到的8株酱油生产菌进行RAPD分析。实验筛选到6个扩增产物谱带多、特征好、覆盖面广的引物:Primer1、Primer5、Primer6、Primer7、Primer8、Primer9,重复实验证明其RAPD-PCR扩增图谱具有较好的稳定性,扩增产物谱带一般4~8条,各实验菌株主带1~4条,次带丰富。根据RAPD-PCR扩增图谱构建的系统进化树较好地吻合了传统的形态分类学,证实了RAPD分子标记在酱油生产菌系统发育分析中应用的可行性。     

水稻品种对美国稻瘟病小种IB-33、IB-45和IE-1的抗性遗传

1995年10月至1997年11月,在美国阿肯色大学水稻研究推广中心,用水稻品种LA110和Jasmine-85与水稻品种Teqing、Katy、Mars、LaGrue和Newbonnet进行不完全双列杂交,对其杂交后代和亲本用美国3个主要稻瘟病菌小种(以下简称小种)IB-33、IB-45和IE-1进行接种鉴定和遗传分析研究.结果表明:亲本LA110、Jasmine-85、Teqing抗所有3个小种.Katy抗小种IB-45和IE-1,感小种IB-33.Mars抗小种IE-1,感小种IB-33和IB-45.LaGrue感所有3个小种.Newbonnet抗小种IB-45,感小种IB-33和IE-1.所有抗病亲本的抗病基因,其F1分别对相应小种呈现显性抗病性.抗病亲本杂交,LA110与Jasmine-85对小种IB-33,LA110与Teqing、Jasmine-85对小种IE-1,及Jasmine-85与Teqing对小种IE-1,是等位的抗病基因.LA110与Teqing对小种IB-33,及Jasmine-85与Teqing对小种IB-33,分别存在三对独立遗传的显性抗病基因.LA110与Teqing、Katy、Newbonnet、Jasmine-85对小种IB-45,Jasmine-85与Teqing、Katy、Newbonnet对小种IB-45,LA110与Katy、Mars对小种IE-1,Jasmine-85与Katy、Mars对小种IE-1,分别存在两对独立遗传的显性抗性基因.抗病亲本LA110或Jasmine-85与感病亲本Mars对小种IB-33,抗病亲本LA110与感病亲本Mars对小种IB-45,具有两对显性互补抗病基因,当两对显性抗病基因同时存在时,表现出抗性.抗病亲本LA110或Jasmine-85与感病亲本Katy、LaGrue、Newbonnet对小种IB-33,抗病亲本LA110与感病亲本LaGrue对小种IB-45,抗病亲本Jasmine-85与感病亲本Mars、LaGrue对小种IB-45,抗病亲本LA110或Jasmine-85与感病亲本LaGrue、Newbonnet对小种IE-1,分别存在一对显性抗病基因.两个亲本正、反交的遗传表现一致.本文也讨论了LA110、Teqing和Jasmine-85三个抗病品种在美国水稻抗病育种中利用的可能性.     

Phenotyping of Partial Physiological Resistance to Rice Sheath Blight

Sheath blight, caused by Rhizoctonia solani, is one of the most important rice diseases worldwide especially under irrigated agro‐ecosystems. To date, no rice accession with complete resistance to sheath blight has been reported. However, a number of genotypes with varying levels of resistance have been reported. Twelve genotypes (including mega varieties) viz. Tetep, Jasmine 85, Te‐Qing, Duduruchi, Betichikon, Khatochalani, D‐6766, D‐256, Swarna, Sarju‐52, MTU‐1010 and Samba Mashuri were evaluated for quantitative measurement of partial physiological resistance to sheath blight under controlled conditions using detached tiller method. Three independent experiments, each involving three replications, were conducted. Seven days after inoculation, the following disease variables were measured: number of lesions, lesion length, vertical sheath colonization (VSC) on the tiller, disease severity, relative vertical sheath colonization (RVSC) and survival of the leaf blade. Variation between rice genotypes was observed for all the disease variables. Disease severity and VSC were the two most correlated variables, whereas the number of lesions and mean lesion length were the least correlated variables. The ranking of varieties often differed depending on the disease variable considered. Amongst the genotypes tested, D‐256, Tetep and Jasmin‐85 had the lowest number of lesions and disease severity. Similarly, Tetep and D‐256 showed the lowest levels of RVSC, whilst Jasmine‐85 was found to be intermediate. D‐6766, Samba Mashuri and Betichikon showed the highest levels of disease variables. The fraction of dead leaves ranged from 0.00 to 0.38. No dead leaves were observed in Te‐Qing, Swarna and MTU‐1010. The highest fraction of dead leaves was observed for Betichikon (0.38) followed by Duduruchi and D‐6766 (0.33). Our results suggest that this method in combination with other phenotyping methods could be used to quantify partial resistance to rice sheath blight.     

Inactivation of an aminoaldehyde dehydrogenase is responsible for fragrance in rice

Rice (Oryza sativa) has two betaine aldehyde dehydrogenase homologs, BAD1 and BAD2, encoded on chromosome four and chromosome eight respectively. BAD2 is responsible for the characteristic aroma of fragrant rice. Complementary DNA clones of both BAD1 and BAD2 were isolated and expressed in E. coli. BAD2 had optimum activity at pH 10, little to no affinity towards N-acetyl-gamma-aminobutyraldehyde (NAGABald) with a Km of approximately 10 mM and moderate affinity towards gamma-guanidinobutyraldehyde (GGBald) and betaine aldehyde (bet-ald) with Km values of approximately 260 microM and 63 microM respectively. A lower Km of approximately 9 microM was observed with gamma-aminobutyraldehyde (GABald), suggesting BAD2 has a higher affinity towards this substate in vivo. The enzyme encoded on chromosome four, BAD1, had optimum activity at pH 9.5, showed little to no affinity towards bet-ald with a Km of 3 mM and had moderate affinity towards GGBald, NAGABald and GABald with Km values of approximately 545, 420 and 497 microM respectively. BAD1 had a half life roughly double that of BAD2. We discuss the implications of these findings on the pathway of fragrance generation in Basmati and Jasmine rice and the potential of rice to accumulate the osmoprotectant glycine betaine.     

Cloning and Characterization of AKR4C14, a Rice Aldo–Keto Reductase, from Thai Jasmine Rice

Aldo–keto reductase (AKR) is an enzyme superfamily whose members are involved in the metabolism of aldehydes/ketones. The AKR4 subfamily C (AKR4C) is a group of aldo–keto reductases that are found in plants. Some AKR4C(s) in dicot plants are capable of metabolizing reactive aldehydes whereas, such activities have not been reported for AKR4C(s) from monocot species. In this study, we have screened Indica rice genome for genes with significant homology to dicot AKR4C(s) and identified a cluster of putative AKR4C(s) located on the Indica rice chromosome I. The genes including OsI_04426, OsI_04428 and OsI_04429 were successfully cloned and sequenced by qRT-PCR from leaves of Thai Jasmine rice (KDML105). OsI_04428, later named AKR4C14, was chosen for further studies because it shares highest homology to the dicot AKR4C(s). The bacterially expressed recombinant protein of AKR4C14 was successfully produced as a MBP fusion protein and his-tagged protein. The recombinant AKR4C14 were capable of metabolizing sugars and reactive aldehydes i.e. methylglyoxal, a toxic by-product of the glycolysis pathway, glutaraldehyde, and trans-2-hexenal, a natural reactive 2-alkenal. AKR4C14 was highly expressed in green tissues, i.e. leaf sheets and stems, whereas flowers and roots had a significantly lower level of expression. These findings indicated that monocot AKR4C(s) can metabolize reactive aldehydes like the dicot AKR4C(s) and possibly play a role in detoxification mechanism of reactive aldehydes.     

A fibronectin-binding protein from rice bran with cell adhesion activity for animal tumor cells.

A rice bran 57-kDa protein was isolated by affinity chromatography with fibronectin immobilized on agarose. This fibronectin-binding protein designated as RB-57 had an amino-terminal amino acid sequence identical with that of a putative mature form of rice hydroxyproline-rich glycoprotein. A distinct feature of the amino acid composition of RB-57 was the high contents of hydroxyproline and proline representing about 45% of the total amino acids. The sugar analysis indicated that arabinose represented 46.8% of the total carbohydrates. RB-57 showed cell adhesion activity for murine Lewis lung carcinoma cells. The result suggests that RB-57 may play a role in plant cell adhesion, although cell adhesion-promoting activity for plant cells remains to be tested.     

Multiple forms of ADPglucose pyrophosphorylase of rice endosperm

ADPglucose pyrophosphorylase from developing rice ( Oryza sativa ) endosperm was purified. The final preparation yielded 6 major protein spots as separated by two-dimensional polyacrylamide electrophoresis. All 6 polypeptides had similar molecular weights of ca 50 kDa and cross-reacted with polyclonal antibodies raised against two main protein bands among them. The results suggest that the rice endosperm ADPglucose pyrophorsphorylase is tetrameric and composed of multiple subunits with similar amino acid structure.     

Development of Desiccation Tolerance during Embryogenesis in Rice (Oryza sativa) and Wild Rice (Zizania palustris) (Dehydrin Expression,Abscisic Acid Content,and Sucrose Accumulation)

The ability of seeds to withstand desiccation develops during embryogenesis and differs considerably among species. Paddy rice (Oryza sativa L.) grains readily survive dehydration to as low as 2% water content, whereas North American wild rice (Zizania palustris var interior [Fasset] Dore) grains are not tolerant of water contents below 6% and are sensitive to drying and imbibition conditions. During embryogenesis, dehydrin proteins, abscisic acid (ABA), and saccharides are synthesized, and all have been implicated in the development of desiccation tolerance. We examined the accumulation patterns of dehydrin protein, ABA, and soluble saccharides (sucrose and oligosaccharides) of rice embryos and wild rice axes in relation to the development of desiccation tolerance during embryogenesis. Dehydrin protein was detected immunologically with an antibody raised against a conserved dehydrin amino acid sequence. Both rice and wild rice embryos accumulated a 21-kD dehydrin protein during development, and an immunologically related 38-kD protein accumulated similarly in rice. Dehydrin protein synthesis was detected before desiccation tolerance had developed in both rice embryos and wild rice axes. However, the major accumulation of dehydrin occurred after most seeds of both species had become desiccation tolerant. ABA accumulated in wild rice axes to about twice the amount present in rice embryos. There were no obvious relationships between ABA and the temporal expression patterns of dehydrin protein in either rice or wild rice. Wild rice axes accumulated about twice as much sucrose as rice embryos. Oligosaccharides were present at only about one-tenth of the maximum sucrose concentrations in both rice and wild rice. We conclude that the desiccation sensitivity displayed by wild rice grains is not due to an inability to synthesize dehydrin proteins, ABA, or soluble carbohydrates.     

Seasonal expression of Bt proteins in transgenic rice lines and the resistance against Asiatic rice borer Chilo suppressalis (Walker)

Laboratory bioassays and field surveys were carried out to compare the resistance of three transgenic rice (Oryza sativa L.) lines including Bt-DL expressing a single gene cry1Ab, Bt-KF6 expressing stacked genes cry1Ac and CpTI genes and Bt-SY63 expressing a fusion gene cry1Ab/cry1Ac, respectively, to an important rice pest Chilo suppressalis (Walker). In addition, enzyme-linked immunosorbent assays (ELISA) were conducted to monitor the Bt protein expressions in rice leaves and stems at different rice growth stages. Results showed that all the transgenic rice lines exhibited significantly high resistance to the pest compared with their corresponding nontransformed isolines. Among the transgenic rice lines, Bt-SY63 and Bt-KF6 had higher resistance to C. suppressalis at early growth stage, but lower resistance at late stages, while the pest resistance of Bt-DL was relatively stable throughout the growing season. The results were consistent with ELISA results showing that Bt protein levels in Bt-SY63 or Bt-KF6 leaves decreased in late growth stages, but were relatively stable in Bt-DL at all growth stages. This demonstrates that the resistance to a pest by Bt plants is positively correlated with Cry protein expression levels in plant tissues. Compared with Bt-SY63 and Bt-KF6, the Bt protein expression levels were significantly lower in Bt-DL, while its resistance to C. suppressalis was the highest. This may suggest that C. suppressalis is more susceptible to Cry1Ab than to Cry1Ac. The data from the current study are valuable for decision-making for commercial use of Bt rice lines and development of appropriate pest control and resistance management strategies for the transgenic rice lines.     

Genomewide comparative phylogenetic and molecular evolutionary analysis of tubby-like protein family in Arabidopsis, rice, and poplar

Tubby-like proteins, which are characterized by a highly conserved tubby domain, play an important role in the maintenance and function of neuronal cells during postdifferentiation and development in mammals. In additional to the tubby domain, most tubby-like proteins in plants also possess an F-box domain. Plants also appear to harbor a large number of TLP genes. To gain insight into how TLP genes evolved in plants, we conducted a comparative phylogenetic and molecular evolutionary analysis of the tubby-like protein gene family in Arabidopsis, rice, and poplar. Genomewide screening identified 11 TLP genes in Arabidopsis, 14 in rice, and 11 in poplar. Phylogenetic trees, domain organizations, and intron/exon structures classified this family into three subfamilies and indicated that species-specific expansion contributed to the evolution of this family in plants. We determined that in rice and poplar, the tubby-like protein family had expanded mainly through segmental duplication events. Tissue-specific expression analysis indicated that functional diversification of the duplicated TLP genes was a major feature of long-term evolution. Our results also demonstrated that the tubby and F-box domains had co-evolved during the evolution of proteins containing both domains.