Endopeptidase polymorphism and linkage of the Ep-D1c null allele with the Lrl9 leaf-rust-resistance gene in hexaploid wheat

The aim of this study was to test whether the null allele Ep-D1c of the endopeptidase Ep-D1 can be used as a marker for the Lrl9 leaf rust resistance gene. The frequency of Ep-D1c was determined in 1134 winter wheat, spring wheat and spelt breeding lines and varieties. Only eight lines were found to carry Ep-D1c. Six of these lines originated from crosses with RL6040, the gene donor for Lrl9. The other two lines were leaf-rust susceptible in the seedling stage and therefore did not carry Lr19. The genetic distance between Ep-D1c and Lr19 was determined in a reciprocal cross between the lines FAP75184 (Ep-D1c, Lr19) and FAP75106 (Ep-D1a, leaf-rust susceptible in the seedling stage). Out of 840 F₂ seedlings screened, 162 were homozygous for Ep-D1c. From 150 of these F₂ plants, F₃ seedlings were screened for segregation for leaf-rust resistance with isolates avirulent on Lr19. Only one F₂ plant produced susceptible F₃ progeny indicating a recombination event between Ep-D1c and Lr19. From these data, a genetic distance of 0.33 ± 0.33cM between Ep-D1c and Lr19 was calculated. The results show that Ep-D1c is a useful marker for a practical breeding programme allowing the rapid identification of plants homozygous for Lrl9.     

Effects of porcine brain hydrolysate on impairment of cognitive learning ability in amyloid β(1–40)‐infused rats

This study assessed whether administering porcine brain hydrolysate (PBH) ameliorates the impairment of spatial cognition learning ability in amyloid β (Aβ)‐infused rats. PBH was prepared using organic solvents (i.e., acetone and ethanol). Enzyme hydrolysates were derived from these PBH and the sequence of the Aβ peptide for infusion was selected. The results indicated the PBH, in particular EP (porcine brain extract with ethanol and protease N), demonstrated the potentials to reduce damage of neurodegenerative disorders in vitro and in vivo. The principal findings of this study indicate that PBH has prolyl endopeptidase inhibitory activity in vitro. Moreover, administering EP to Aβ(1–40)‐infused rats significantly improves their performance on reference, spatial performance, and working memory tests during water maze tasks; concurrent proportional decreases are also observed in malondialdehyde levels, acetylcholinesterase (AChE) activity, and Aβ accumulation levels in brain tissues. The PBH was suggested to ameliorate learning deficits associated with Alzheimer's disease by inhibition of lipid peroxidation in the brain of Aβ infused rat.     

Linkage relations among eyespot resistance gene Pch2, endopeptidase Ep-A1b, and RFLP marker Xpsr121 on chromosome 7A of wheat

Marker-based selection of Ep-D1b has been used successfully to incorporate Pch1, the gene for eyespot resistance on chromosome 7D, into commercial wheat. However, attempts to transfer resistance conferred by Pch1 (on chromosome 7A) through selection for Ep-A1b have not always been successful. Linkage relations among eyespot resistance gene Pch2, a gene encoding for an isozyme of endopeptidase, Ep-A1b, and RFLP marker Xpsr121 on chromosome 7A were determined using 80 homozygous recombinant substitution lines. The recombinant lines were derived from eyespot susceptible ‘Chinese Spring’ hybridized with a resistant disomic substitution line of ‘Cappelle Desprez’ that has chromosome 7A substituted into ‘Chinese Spring’. Segregations of Pch2, Ep-A1b and Xpsr121 fit an expected 1:1 single-locus ratios based on χ² tests. Linkage analysis revealed that Pch2 was not tightly linked to Ep-Alb (15% recombination). However, close linkage (3.8% recombination) existed between Ep-A1b and Xpsr121. The order of these loci is Pch2-Xpsr121-Ep-A1b. Unlike Pch1 and Ep-D1b, where little or no recombination is found, Pch1 and Ep-A1b showed considerable recombination and therefore linkage cannot be utilized efficiently in marker-based selection.     

Amplified fragment length polymorphism-derived microsatellite sequence linked to the Pch1 and Ep-D1 loci in common wheat

Amplified fragment length polymorphism (AFLP) markers linked to the Aegilops ventricosa‐derived chromosome segment in ‘VPM1’ on which the eyespot resistance gene, Pch1, and the endopeptidase gene, Ep‐D1b, occur were identified. One marker was isolated from the gel, cloned and sequenced. Sequence analysis revealed a microsatellite repeat motif. Sequence‐specific primers were designed to amplify a product containing the repeat motif, and the microsatellite marker was tested for cosegregation with the Ep‐D1b allele. Distinct alleles were produced by the Pch1 sources, normal wheat and wheat containing the Lr19 translocation. A recombination frequency of 0.02 was calculated between the microsatellite marker and Ep‐D1.     

水稻叶片光合功能衰退过程中内肽酶活力的变化

以两个粳稻品种武育粳3号和97-7作材料研究了光合功能衰退进程中叶片中可溶性蛋白质、游离氨基酸和内肽酶活力的变化。武育粳3号剑叶的可溶性蛋白质含量比97-7高,但其含量下降比97-7快。武育粳3号的剑叶和第13叶在光合功能衰老转折点到来之前10 d左右游离氨基酸含量开始上升,而97-7则在转折点之前1～2 d才开始上升。在光合功能衰退的可逆阶段,内肽酶的活力维持在较低水平,只是在可逆阶段的后期内肽酶的活力有轻微的升高。但进入不可逆阶段后,内肽酶的活力则急剧升高,并达到很高的水平。武育粳3号的第13 叶和剑叶中的内肽酶活力变化基本上是平行的,而97-7的第13叶内肽酶的活力急剧上升期却迟于剑叶。这表明,与武育粳3号相比,在97-7叶片衰老过程中,叶片中的蛋白质降解、转化为可输出氮的进程缓慢。这可能是97-7结实率不高、产量潜力不能充分发挥的一个重要原因。     

Endopeptidase EP-1 as a Marker for the Eyespot Resistance Gene Pch-1 from Aegilops ventricosa in Wheat Line 'H-93-70'

Biochemical marker endopeptidase EP-1 was used to determine that an eyespot resistance gene, transferred from Aegilops ventricosa to the hexaploid wheat line ‘H-93-70’ is located on the long arm of the chromosome 7D.     

黄瓜叶片生长期间内肽酶活性和同工酶变化及其生化特性

研究了黄瓜自然生长期间叶片可溶性蛋白质降解、内肽酶活性及其同工酶的变化,并对叶片衰老后期粗提液内肽酶的基本生化特性进行了研究.结果表明,黄瓜叶片内肽酶活性在叶片伸出5 d时较低,叶片衰老后期(叶片伸出35 d)显著升高,Rubisco蛋白发生明显降解;叶片全展以后叶片蛋白质含量与内肽酶活性显著负相关.黄瓜叶片粗提液总内肽酶的最适pH为7.0,最适温度为40 ℃,内肽酶活性可被Ca2 、Zn2 、半胱氨酸等激活.内肽酶抑制剂试验显示,丝氨酸类型的内肽酶活性占内肽酶总活性的57%左右.采用以明胶为底物的内肽酶梯度凝胶电泳活性染色方法,检测到叶片中至少有4种内肽酶同工酶,各同工酶在叶片生长过程中出现的时间和活性大小不同,丝氨酸型内肽酶同工酶活性随叶片生长明显增强,暗示该类内肽酶在黄瓜叶片衰老过程中可能具有重要作用.     

Changes in Endopeptidase Activity during Photosynthetic Declination in Rice Leaf

Two japonica rice varieties, Wuyujing 3 and 97-7, were used to study the changes in contents of soluble protein, free amino acids and endopeptidase activity during photosynthetic declination. The content of soluble protein in flag leaf of cv. Wuyujing 3 was higher than that of cv. 97-7, but decreased rapidly in Wuyujing 3. Free amino acids in flag leaf and the thirteenth leaf of Wuyujing 3 started to increase 10 days before the turning point of photosynthetic declination (TPPD), while it occurred just 1-2 days before TPPD in the flag leaf and the thirteenth leaf of 97-7. During reversible phase of photosynthetic declination, endopeptidase activity remained at a low level and increased slightly only in the later part of this phase. Then it rose up rapidly at irreversible decline phase and reached a very high level. For Wuyujing 3, the change in endopeptidase activity in the thirteenth leaf was parallel to that in flag leaf. However, for 97-7, the rapid increase of endopeptidase activity in the thirteenth leaf started later than that of flag leaf. The results implied that the rate of protein breakdown and conversion to transportable nitrogen in leaves of 97-7 was slower than that in leaves of Wuyujing 3 during photosynthetic declination and it led to relatively lower seed setting rate and fully filling grains rate in 97-7. This may be one of the important reasons why 97-7 could not bring the high yield potentiality into play and the findings may be taken into consideration while breeding for high potential varieties in future.     

Structure,chemical composition and enzymatic activities of pearlings and bran obtained from pearled wheat (Triticum aestivum L.) by roller milling

While abrasive pearling (also referred to as debranning) of wheat kernels prior to milling increases the quality of the resultant flour for producing bread, the potential applications of the co-products of pearling is largely unknown. We studied the impact of different degrees of pearling (0, 3, 6, 9 and 12% by weight) on the composition of pearlings and bran obtained when subsequently roller milling pearled wheat kernels. Pearling does not remove the kernel outer tissues homogeneously as abrasion affects especially the accessible parts of the kernels. Nevertheless, the first 3% removed consisted of mainly pericarp. With 6% or more removed, a significant amount of starchy endosperm ended up in the pearlings. The starting bran material and bran obtained by subsequent roller milling of pearled wheat kernels had similar compositions but the latter had a lower average particle size. Moreover, removal of the outermost kernel layers substantially decreased the enzyme activity levels in the bran.     

铝胁迫与小麦叶片的内肽酶活性及活性氧的产生

采用温室内自然光下小麦种子培养实验 ,研究了Al3 胁迫下和H2O2 处理时小麦幼苗叶片中的内肽酶活性和可溶性蛋白质的含量变化。结果表明 ,Al3 的胁迫和H2O2 处理均使小麦幼苗叶片中的内肽酶活性升高 ,可溶性蛋白质含量下降。Al3 的胁迫下 ,H 2O2 的清除剂ASA(抗坏血酸 )的加入使内肽酶活性明显下降 ,可溶性蛋白质的含量明显增加。Al3 胁迫下小麦幼苗叶片中的内肽酶活性升高和可溶性蛋白质含量下降的原因之一是产生了活性氧—H2O2。