Action de la proline exogène sur l'activité de la voie du glycolate chez Nicotiana tabacum cv. Xanthi n.c.

The effect of exogenous proline on the activity of the glycolate pathway in Nicotiana tabacum cv. Xanthi n.c. An exogenous proline supply in the light provokes an increase in free glycine concentration in apical tissues or in leaf disks of vegetative Nicotiana tabacum L. cv. Xanthi n.c. This does not occur in the equivalent tissues of tobacco plants after floral induction, these being naturally rich in proline. In vegetative tobacco, we have tried to determine this specific action of exogenous proline. With ¹⁴C glycine, ¹⁴CO₂ experiments (Pulse-chase) and glycine decarboxylase activity determinations, we observed that glycine-serine transformation was inhibited by proline supply. Presently it is important to determine if endogenous proline acts on the same reaction.     

Indices of water deficit in susceptible and resistant cucumbers in response to infection by cucumber mosaic virus

Indices of water deficit were determined under conditions of non-limited water supply in cucumber mosaic virus (CMV)-infected seedlings of the susceptible cv. Bet Alpha. An increase in the concentration of soluble solids, decrease of water and osmotic potentials, and increase of proline concentration were found in the CMV-infected cotyledons. In the cv. Shimshon, which is resistant to CMV, virus infection caused only a slight change in the concentration of the soluble solids and in the osmotic potential; water potential and proline content were not affected. Concomitantly, infectivity of cotyledons by CMV was much lower in the tolerant cv. Shimshon than in the susceptible cv. Bet Alpha. The possible association of water deficit with virus-induced growth retardation is discussed.     

Three proline rotamases involved in calcium homeostasis play differential roles in stress tolerance,virulence and calcineurin regulation of Beauveria bassiana

FK506‐sensitive proline rotamases (FPRs), also known as FK506‐binding proteins (FKBPs), can mediate immunosuppressive drug resistance in budding yeast but their physiological roles in filamentous fungi remain opaque. Here, we report that three FPRs (cytosolic/nuclear 12.15‐kD Fpr1, membrane‐associated 14.78‐kD Fpr2 and nuclear 50.43‐kD Fpr3) are all equally essential for cellular Ca²⁺ homeostasis and contribute significantly to calcineurin activity at different levels in the insect‐pathogenic fungus Beauveria bassiana although the deletion of fpr1 alone conferred resistance to FK506. Radial growth, conidiation, conidial viability and virulence were less compromised in the absence of fpr1 or fpr2 than in the absence of fpr3, which abolished almost all growth on scant media and reduced growth moderately on rich media. The Δfpr3 mutant was more sensitive to Na⁺, K⁺, Mn²⁺, Ca²⁺, Cu²⁺, metal chelate, heat shock and UVB irradiation than was Δfpr2 while both mutants were equally sensitive to Zn²⁺, Mg²⁺, Fe²⁺, H₂O₂ and cell wall‐perturbing agents. In contrast, the Δfpr1 mutant was less sensitive to fewer stress cues. Most of 32 examined genes involved in DNA damage repair, Na⁺/K⁺ detoxification or osmotolerance and Ca²⁺ homeostasis were downregulated sharply in Δfpr2 and Δfpr3 but rarely so affected in Δfpr1, coinciding well with their phenotypic changes. These findings uncover important, but differential, roles of three FPRs in the fungal adaptation to insect host and environment and provide novel insight into their essential roles in calcium signalling pathway.     

内生细菌对盐胁迫下小麦幼苗脯氨酸和丙二醛的影响

以周麦18为试验材料,以内生菌252和254为供试菌株,试验采用盆栽法设置盐胁迫组、盐胁迫接菌组和对照组,研究不同盐浓度胁迫下接菌处理对小麦幼苗脯氨酸(Pro,proline)和丙二醛(MDA,malondialdehyde)含量的影响。结果表明:小麦幼苗在盐胁迫环境下,随着培养时间和盐浓度增加,盐胁迫组Pro含量先升后降。在前中期,盐胁迫组小麦Pro含量逐渐积累,提高了小麦抗逆性;在后期Pro含量较中期有一定程度下降,小麦逐渐对盐环境产生适应性。随着盐浓度升高,盐胁迫组小麦在不同培养阶段对盐环境的抵抗能力不同,每个时期对盐抵抗能力有一阈值,超过此盐浓度Pro含量下降,抵抗能力减弱。接菌处理后,小麦Pro含量升高,提高了抗盐胁迫能力,第14天时修复效果最明显,接菌252处理组和254处理组分别在盐浓度为200 mmol/L和250 mmol/L时高出盐胁迫组133.48%和91.48%。盐胁迫组MDA含量随时间延长先降后升,与Pro含量变化趋势相反。在前中期,盐胁迫组小麦膜质过氧化程度不高,对小麦正常代谢活动尚未造成严重影响,在后期MDA含量升高,盐胁迫组小麦逐渐枯萎。接菌后,降低了幼苗细胞膜脂化程度,提高了小麦存活率,促进幼苗生长,MDA含量在21 d时均低于盐胁迫组,修复效果显著。     

黄河三角洲河滩与潮滩芦苇对盐胁迫的生理生态响应

研究黄河三角洲两种生境芦苇对盐胁迫的生理生态响应差异,能为退化滨海湿地生态修复中芦苇植株来源的选择提供重要的理论支持。在盐胁迫(300 mmol/L NaCl)下,比较研究了黄河三角洲河滩芦苇(低盐生境)和潮滩芦苇(高盐生境)叶片中的Na~+含量、根部分生区Na~+流速、叶片的光合作用参数、H_2O_2的含量、抗氧化酶的活性、丙二醛和脯氨酸的含量。结果表明:盐胁迫显著提高了河滩芦苇叶片中Na~+含量,但对潮滩芦苇叶片Na~+影响不显著。进一步通过非损伤微测技术研究发现,盐胁迫后,潮滩芦苇比河滩芦苇的根部分生区的Na~+外排流速更高(潮滩芦苇:(1982.05±122.74) pmol cm~(-2) s~(-1)vs.(87.93±12.94) pmol cm~(-2) s~(-1),P0.01;河滩芦苇:(1574.16±458.90) pmol cm~(-2) s~(-1)vs.(-126.88±23.01) pmol cm~(-2) s~(-1),P0.01),能有效调节细胞内离子平衡。另外,盐处理7天后潮滩芦苇光合速率((16.36±1.09)μmol m~(-2) s~(-1)vs.(22.79±0.67)μmol m~(-2) s~(-1),P0.01)显著高于河滩芦苇((12.71±0.97)μmol m~(-2) s~(-1)vs.(23.81±0.55)μmol m~(-2) s~(-1),P0.01)。盐胁迫诱导了两种芦苇叶片中H_2O_2含量和丙二醛含量的显著升高,而脯氨酸及抗氧化酶的活性也随之显著升高,并且也显著升高了潮滩芦苇的谷胱甘肽还原酶(GR)活性((6.90±1.73) U/mg蛋白质vs.(3.54±0.54) U/mg蛋白质,P0.05)。综上,潮滩芦苇比河滩芦苇更适应盐胁迫,因其根系具有更高的排Na能力,且脯氨酸含量及抗氧化酶活性较高,提高了抗逆性。因此,在黄河三角洲退化滨海湿地生态修复中,可以优先选择潮滩芦苇作为植被修复的材料。     

Genetic analysis of Pycr1 and Pycr2 in mice

The final step in proline biosynthesis is catalyzed by three pyrroline-5-carboxylate reductases, PYCR1, PYCR2, and PYCR3, which convert pyrroline-5-carboxylate (P5C) to proline. Mutations in human PYCR1 and ALDH18A1 (P5C Synthetase) cause Cutis Laxa (CL), whereas mutations in PYCR2 cause hypomyelinating leukodystrophy 10 (HLD10). Here, we investigated the genetics of Pycr1 and Pycr2 in mice. A null allele of Pycr1 did not show integument or CL-related phenotypes. We also studied a novel chemically-induced mutation in Pycr2. Mice with recessive loss-of-function mutations in Pycr2 showed phenotypes consistent with neurological and neuromuscular disorders, including weight loss, kyphosis, and hind-limb clasping. The peripheral nervous system was largely unaffected, with only mild axonal atrophy in peripheral nerves. A severe loss of subcutaneous fat in Pycr2 mutant mice is reminiscent of a CL-like phenotype, but primary features such as elastin abnormalities were not observed. Aged Pycr2 mutant mice had reduced white blood cell counts and altered lipid metabolism, suggesting a generalized metabolic disorder. PYCR1 and -2 have similar enzymatic and cellular activities, and consistent with previous studies, both were localized in the mitochondria in fibroblasts. Both PYCR1 and -2 were able to complement the loss of Pro3, the yeast enzyme that converts P5C to proline, confirming their activity as P5C reductases. In mice, Pycr1; Pycr2 double mutants were sub-viable and unhealthy compared to either single mutant, indicating the genes are largely functionally redundant. Proline levels were not reduced, and precursors were not increased in serum from Pycr2 mutant mice or in lysates from skin fibroblast cultures, but placing Pycr2 mutant mice on a proline-free diet worsened the phenotype. Thus, Pycr1 and -2 have redundant functions in proline biosynthesis, and their loss makes proline a semi-essential amino acid. These findings have implications for understanding the genetics of CL and HLD10, and for modeling these disorders in mice.     

螺旋藻在光胁迫时的抗逆性研究

螺旋藻对于环境的变化有很强的适应性.以钝顶螺旋藻为实验材料,测定螺旋藻在受到较强光照胁迫时藻体的电导率以及脯氨酸、丙二醛和过氧化氢酶系的含量.在3000 lx光照下,螺旋藻6个藻株的电导率以及脯氨酸、丙二醛和过氧化氢酶系的含量比在1000 lx光照下明显升高,螺旋藻的电导率最高上升了2～6倍;细胞内的脯氨酸含量最多增加5倍,最少增加13%;丙二醛的含量增加40%～100%;过氧化氢酶的含量上升范围在19%～80%,过氧化物酶的含量上升范围在20%～100%.说明螺旋藻在受到光胁迫时自身会启动相关保护机制,产生一定的抗逆性,以适应环境的变化.     

<i>Trichoderma</i>-Induced Improvement in Growth,Photosynthetic Pigments,Proline, and Glutathione Levels in <i>Cucurbita pepo</i> Seedlings under Salt Stress

Salt stress is one of the major abiotic stress in plants. However, traditional approaches are not always efficient in conferring salt tolerance. Experiments were conducted to understand the role of Trichoderma spp. (T. harzianum and T. viride) in growth, chlorophyll (Chl) synthesis, and proline accumulation of C. pepo exposed to salinity stress. There were three salt stress (50, 100, and 150 mM NaCl) lavels and three different Trichoderma inoculation viz. T. harzianum, T. viride, and T. harzianum + T. viride. Salt stress significantly declined the growth in terms of the shoot and root lengths; however, it was improved by the inoculation of Trichoderma spp. C. pepo inoculated with Trichoderma exhibited increased synthesis of pigments like chl a, chl b, carotenoids, and anthocyanins under normal conditions. It was interesting to observe that such positive effects were maintained under salt-stressed conditions, as reflected by the amelioration of the salinity-mediated decline in growth, physiology and antioxidant defense. The inoculation of Trichoderma spp. enhanced the synthesis of proline, glutathione, proteins and increased the relative water content. In addition, Trichoderma inoculation increased membrane stability and reduced the generation of hydrogen peroxide. Therefore, Trichoderma spp. can be exploited either individually or in combination to enhance the growth and physiology of C. pepo under saline conditions.     

A CRYGC gene mutation associated with autosomal dominant pulverulent cataract

Purpose

To describe at molecular level a family with pulverulent congenital cataract associated with a CRYGC gene mutation.

Methods

One family with several affected members with pulverulent congenital cataract and 230 healthy controls were examined. Genomic DNA from leukocytes was isolated to analyze the CRYGA-D cluster, CX46, CX50 and MIP genes through high-resolution melting curve and DNA sequencing.

Results

DNA sequencing in the affected members revealed the c.143G>A mutation (p.R48H) in exon 2 of the CRYGC gene; 230 healthy controls and ten healthy relatives were also analyzed and none of them showed the c.143G>A mutation. No other polymorphisms or mutations were found to be present.

Conclusion

In the present study, we described a family with pulverulent congenital cataract that segregated the c.143G>A mutation (p.R48H) in the CRYGC gene. A few mutations have been described in the CRYGC gene in autosomal dominant cataract, none of them with pulverulent cataract making clear the clinical heterogeneity of congenital cataract. This mutation has been associated with the phenotype of congenital cataract but also is considered an SNP in the NCBI data base. Our data and previous report suggest that p.R48H could be a disease-causing mutation and not an SNP.