The Separation And Purification Of Potato Endogenous Bacteria And Influence The Growth And Development Of Potato

Endophytic fungi is a class of special habitat microbial resources, widelydistributed in the moss, ferns, gymnosperms, and angiosperms, with long-termcooperative coevolution, they can produce the same or similar compounds with theirhost, and most compounds have novel structure. In this thesis, the resources of thegroup cultivates the seeding of potato LK99(Solanum tuberosum L. cv. LK99),cultivated the seedling endogenous fungus and the endogenous fungus to the potatoLK99group cultivates the seedling growth to its potato LK99group the influence toconduct the preliminary study, The results were as follows:1. Taking grows15days of potato LK99group to cultivate the seedling, cuts thelength approximately1.0cm the belt axial bud stem section, vaccinates in the potatosucrose agar-agar culture medium (PSA) and on the MS culture medium, under light(2200Lx,16hr light/8hr darkness,28℃) and28℃dark raise.3days later, under lightand under the dark condition grew on the PSA culture medium group cultivates aroundthe seedling to present the yellow fungus block, but under light and under the darkcondition grew on the MS culture medium group cultivates around the seedling theaseptic block appearance. After the single colony separation, the endogenous fungus'sphysiological biochemistry experiment and16S rDNA fragment's expansion and thesequential analysis knew: Under the light grows on the PSA culture medium groupcultivates the fungus block which around the seedling appears to separate to2kinds offungi, Belonging to Bacillus sp. And Bacillus subtilis, named as Bacillus sp. LW01andBacillus subtilis LS01. Separated into two kinds of bacteria grown in tissue culturesurrounding plaque on PSA medium under dark conditions belong Leifsonia sp. AndBacillus sp. Named Leifsonia sp. DH01and Bacillus sp. DW01.2. Takeing grows15day of potato LK99group to cultivate the seedling, cuts thelength approximately1.0cm the belt axial bud stem section, vaccinates on the MSculture medium, under the light (2200Lx,16hr the light/8hr darkness,28℃) raisesabout5weeks, then cultivates in the bottle toward the group to pour into25mL theliquid MS culture medium, in dark22℃raises about5weeks, treats ties the potato. Theselection diameter is1.5cm microtuber, all are two, section vaccination in potatosucrose agar-agar culture medium (PSA) and on MS culture medium, under light (2200Lx,16hr light/8hr darkness,28℃) and28℃dark raise.3days later, under lightand under the dark condition grew on around the PSA culture medium miniature potatopresented the yellow fungus block, but under light and under the dark condition grew onaround the MS culture medium miniature potato the aseptic block appearance. After thesingle colony separation, the endogenous fungus's physiological biochemistryexperiment and16S rDNA the fragment expansion and the sequential analysis knew:Under the light grows the fungus block which appears on around the PSA culturemedium's miniature potato to separate to2kinds of fungi, Belonging to the Bacillussubtilis and Bacillus sp. Named Bacillus subtilis SLH01and the Bacillus sp. SLB01.Growth under dark conditions at the PSA medium on tissue culture around bacteriablock the isolated bacteria belonging to Bacillus sp., Named as Bacillus sp. SDB01.3. The raise separation obtains7kinds of endogenous fungi, treat the endogenousfungus OD600solstice0.5, cultivates seeding stem the group in the vaccine the immerse,takes out immediately changes over to cultivates the bottle including the MS culturemedium group in28℃,2200Lx,16hr under the light/8hr dark condition to raise. TheDH01strain and the LW01strain have not grown on the PSA culture medium, when the3rd week, the DH01strain and the LW01strain cultivate seeding length to the group tosuppress obviously. The endogenous bacterium DH01treatment group cultivates theseedling, raises after5weeks under the light, the group cultivates the seedling short theroot, sturdy, and the non-lateral root formation, the root color is a vandyck brown, thecontrol group cultivates the seedling tall and slender the root luxuriant, slightly has thegreen, the lateral root to be developed, The entire average fresh weight reduced1.45%compared to the control group, the single average aerial parts fresh weight increased15.63%compared to the control group, the root average fresh weight reduced40.48%compared to the control group. The entire average dry weight, the single average aerialparts dry weight increased6.93%separately compared to the control group,241.30%,the root average dry weight reduced471.43%compared to the control group, the abovedata had indicated that endogenous bacterium DH01had the obvious inhibitory actionto the root, but the biomass accumulation had the obvious promoter action to the placeon. Endogenous bacterium LW01is contaminated the group to cultivate the seedling,raises after5weeks under the light, the group cultivates the seedling the root to be talland slender, slightly has the green, the lateral root to be developed, cultivates theseedling with the control group the root not obvious difference, the entire average freshweight reduced13.9%compared to the control group, the single average aerial partsfresh weight reduced17.9%compared to the control group, the root average fresh weight reduced3.1%compared to the control group. The entire average dry weight, thesingle average aerial parts dry weight increased8.7%separately compared to the controlgroup,8.8%, the root average dry weight increased8.8%compared to the control group,the above data had indicated that endogenous bacterium LW01to biomass accumulationpromoter action, but to fresh weight accumulation inhibitory action. The DW01strainand the LS01strain lower in its OD600to0.18are contaminated the group to cultivatethe seedling, may in1~2days, the DW01strain and the LS01strain on the potatosucrose culture medium the massive multiplication, causes the group to cultivate theseedling death finally.SLH01is contaminated the group to cultivate the seedling, raises5weeks later, itssingle average aerial parts fresh weight increased33.63%compared to the control group,the single average root fresh weight reduced73.78%compared to the control group, thesingle average aerial parts dry weight increased50.85%compared to the control group,the single average root dry weight reduced29.75%compared to the control group. Iscontaminated the group after SDB01to cultivate the seedling, raises5weeks later, itssingle average aerial parts fresh weight increased103.84%compared to the controlgroup, the single average root fresh weight reduced69.80%compared to the controlgroup, the single average aerial parts dry weight increased106.76%compared to thecontrol group, the single average root dry weight reduced62.75%compared to thecontrol group. The SLH01strain and the SDB01strain are contaminated the groupcultivates the seedling5weeks later, its short, sturdy, and the non-lateral root shape, theroot color is a vandyck brown, and is contaminated the spot to expand. But the controlgroup cultivates the seedling tall and slender the root luxuriant, slightly has the green,the lateral root to be developed. The result indicated that the SLH01strain and theSDB01strain have the promoter action to the biomass accumulation, but has theobvious inhibitory action to the root growth, the SLB01strain lowers in its OD600to0.18is contaminated the group to cultivate the seedling, causes the group in2~4days tocultivate the seedling death.