| The Cereal Cyst Nematode (CCN) has caused important crop damage all over the world, and has been one of the major constraints on wheat and barly production. Hatch of H. avenae varies according to region and climate. In order to make clear the responses of H. avenae from Tai’an to environment, experiments were carried out in vitro and field on hatching of second stage larvae (J2) for the Tai’an population of H. avenae, responses of H. avenae to different planting patterns, wheat cultivars’ resistance against H. avenae, and comparison of total protein on diapause and non-diapause H. avenae in2010and2011. The main results as follows:1. Response of CCN to nature and artificial condition changesResponse of eggs in CCN cysts was different from free eggs to the same temperature treatments. The optimum temperature treatment for emergence of J2from cysts of H. avenae was15℃after pre-treated at4℃for30d, then the treatment of15℃constant temperature, and there was no significant difference between the two treatments. J2hatched the most at15℃for free eggs at different temperature treatments and was significantly higher than other temperature treatments, then the treatment of15℃after pre-treated at4℃for30d. There were no hatch peaks at4℃constant temperature treatment for free eggs and eggs in cyst.Hatch of H. avenae was adapted to changes of external temperatures. Cysts in soil were almost empty in April, so it was removed from hatch experiments. Cysts collected from the field treated at15℃had high emergence rate from November to March the next year and were significantly higher than the other months of round year.A little white females produced in April, and number of new cyst reached highest value in August. Number changes of empty cysts in soil were similar to that of total cysts in round year. Numbers of empty and new cysts were high in June and peak appeared in the same month. The number of J2in soil was increased as temperature dropped, reached peak value in March and significantly higher than the leftover11months. In general, number of eggs was higher than that of J2in cysts in one year. There were no J2s in new cysts in April and May. J2s existed in cysts during CCN diapause in summer and were inactive. Number of eggs in cysts was downtrend from October to March next year, while number of J2s has no significant changes.2. Identification of wheat variety resistance to Heterodera avenaThere were no resistance varieties to Heterodera avenae among tested wheat variety (line)., all cultivars were high susceptable (HS). In the present study, there was no relative between growth character of wheat root and numbers of new cysts under the same inoculation number of J2.3. CCN response to different planting paternsThe experiment was the fist year of planting pattern cycle systems, there were no significant difference at decreased number of CCN before nonhost crops seeding and harcestion, so were at two different wheat sowing period. Experiment is being studied in order to make clear CCN response to different planting paterns.4. Comparision of total protein between diapause and non-diapause CCN eggsCCN cell crushed with grinding homogenate method accronding to character of cyst and experimental conditions.7protein extraction reagents were used to extract total protein, and then examined by SDS-PAGE. E (20%sucrose,50mM Tris-HCl (pH7.1),0.5%Triton X-100,0.1%SDS) was selected for further experiment by coparing electrophoretic band and extraction cost between7protein extraction reagents.Differences of total protein between diapause and non-diapaues CCN eggs were examined by SDS-PAGE and compared using reagent E. Total protein content was highest in diapause CCN eggs, and lowest in new CCN eggs at4℃-40d treatment.33electrophoretic bands were examined in diapause CCN eggs, so were in CCN eggs at4℃-40d treatment, while the most electrophoretic bands were examined in treatment of new CCN eggs placed at4℃-40d then transferred to15℃for7days. There was no200KD protein band and existed12.7KD protein spectrum in CCN eggs of treatment at4℃-40d; There was5more protein bands in treatment of CCN eggs placed at4℃-40d then transferred to15℃for7days compared with diapause CCN egg protein.5protein bands were exist in treatment of4℃-40d then transferred to15℃-7d, while none in treatment of4℃-40d.Total protein of CCN eggs was extracted using urea direct extraction solution, then was investigated by two-dimensional electrophoresis (2-DE) and image analysis software ImageMaster M2D Platinum5.0. A total of230protein spots were detected in the2-DE pattern of diapause and non-diapause CCN eggs, of which109belonged to non-diapause eggs and121belonged to diapause eggs. Comparison showed that90pairs were matched in the2-DE patterns of both diapause and non-diapause CCN eggs, with the match percentage as high as74.4%. There are31specific protein spots, of which14protein spots specific to diapause and17specific to non-diapause eggs. |
PDF Full Text Request