The southern root-knot nematode, Meloidogyne incognita, is one of the most prevalent and damaging plant-parasitic nematodes in the world and causes serious damages to agricultural production. We cloned a mitochondrial...The southern root-knot nematode, Meloidogyne incognita, is one of the most prevalent and damaging plant-parasitic nematodes in the world and causes serious damages to agricultural production. We cloned a mitochondrial ATP synthase b subunit gene fragment of M. incognita (MiASB) based on the nematode genomics prediction. By soaking in the MiASB dsRNA solution, the hatching of RNAi treated eggs was reduced by 60% compared to negative control and by 64% compared to untreated control. Mortality of RNAi treated second stage juvenile (J2) was 8.6 times higher than that of negative control and 26 times higher than the untreated control. Inoculating the RNAi treated egg masses and J2 to tomato seedlings showed the pathogencity was significantly reduced. For the RNAi treated egg masses, the amount of root galls on silence treated seedlings was reduced by 92% compared to that on the negative control seedlings, and reduced by 93% compared to that on untreated control seedlings. For the treated J2, the amount of root galls on silence treated seedlings was reduced by 83% and 86% compared to negative and untreated control seedlings, respectively. The study revealed the MiASB silence had a positive effect on prevention and control of root-knot nematode disease, and also showed that the MiASB may be involved in the pathogenesis of nematode, which provided new ideas and ways to the research of nematode pathology and nematode disease control.展开更多
Mitochondrial ATP synthase is responsible for the production of the majority of the cellular ATP, which is composed of two major units: Fo and F1. Although much is known about the active complex (5 subunits (αβ...Mitochondrial ATP synthase is responsible for the production of the majority of the cellular ATP, which is composed of two major units: Fo and F1. Although much is known about the active complex (5 subunits (αβγδε)), the role of the α subunit in the catalytic mechanism remains unclear, particularly in bivalve animals. This study first cloned and identified the full-length sequence of the mitochondrial H^+-ATP synthase α subunit cDNA gene in Pinctada fucata using the reverse transcriptase polymerase chain reaction (RT-PCR) technique. The Pinctada fucata mitochondrial H^+-ATP synthase α subunit contains 1991 nucleotides, with the translation start site at nt 48 (ATG) and the stop codon at nt 1660 (TAA), encoding a polypeptide 553 amino acids in length, which shares high similarity to that of other animals (81% identity to fruit fly, 82% to carp, and 83% to humans). Alignment analysis of the well-conserved amino acid domains in the ATPase α subunit, the call3 signal transduction domain, showed that two residues (Asp^358 and Asn^359) differ from any other ATP synthase α subunit. In situ hybridization analysis was used to reveal the wide-spread distribution of mitochondrial H^+-ATP synthase in various tissues in Pinctada fucata. This work will help further research on pearl energy metabolism to increase the output and quality of pearls to more efficiently utilize our rich pearl oyster resources.展开更多
文摘The southern root-knot nematode, Meloidogyne incognita, is one of the most prevalent and damaging plant-parasitic nematodes in the world and causes serious damages to agricultural production. We cloned a mitochondrial ATP synthase b subunit gene fragment of M. incognita (MiASB) based on the nematode genomics prediction. By soaking in the MiASB dsRNA solution, the hatching of RNAi treated eggs was reduced by 60% compared to negative control and by 64% compared to untreated control. Mortality of RNAi treated second stage juvenile (J2) was 8.6 times higher than that of negative control and 26 times higher than the untreated control. Inoculating the RNAi treated egg masses and J2 to tomato seedlings showed the pathogencity was significantly reduced. For the RNAi treated egg masses, the amount of root galls on silence treated seedlings was reduced by 92% compared to that on the negative control seedlings, and reduced by 93% compared to that on untreated control seedlings. For the treated J2, the amount of root galls on silence treated seedlings was reduced by 83% and 86% compared to negative and untreated control seedlings, respectively. The study revealed the MiASB silence had a positive effect on prevention and control of root-knot nematode disease, and also showed that the MiASB may be involved in the pathogenesis of nematode, which provided new ideas and ways to the research of nematode pathology and nematode disease control.
基金Supported by the National High-Tech Research and Development (863) Program of China (No. 2003AA603430)the National Natural Science Foundation of China (No. 30371092)
文摘Mitochondrial ATP synthase is responsible for the production of the majority of the cellular ATP, which is composed of two major units: Fo and F1. Although much is known about the active complex (5 subunits (αβγδε)), the role of the α subunit in the catalytic mechanism remains unclear, particularly in bivalve animals. This study first cloned and identified the full-length sequence of the mitochondrial H^+-ATP synthase α subunit cDNA gene in Pinctada fucata using the reverse transcriptase polymerase chain reaction (RT-PCR) technique. The Pinctada fucata mitochondrial H^+-ATP synthase α subunit contains 1991 nucleotides, with the translation start site at nt 48 (ATG) and the stop codon at nt 1660 (TAA), encoding a polypeptide 553 amino acids in length, which shares high similarity to that of other animals (81% identity to fruit fly, 82% to carp, and 83% to humans). Alignment analysis of the well-conserved amino acid domains in the ATPase α subunit, the call3 signal transduction domain, showed that two residues (Asp^358 and Asn^359) differ from any other ATP synthase α subunit. In situ hybridization analysis was used to reveal the wide-spread distribution of mitochondrial H^+-ATP synthase in various tissues in Pinctada fucata. This work will help further research on pearl energy metabolism to increase the output and quality of pearls to more efficiently utilize our rich pearl oyster resources.