摘要
目的:通过人工模拟气候室试验,验证冰核细菌是诱发和加重香蕉霜冻害的一个重要因素。通过对冰核细菌的抑菌、杀菌和破坏冰核活性等试验方法筛选出一种低毒、高效防霜冻药剂,建立有效含量测定方法并初步检验其稳定性;通过大田试验检验防霜冻药剂对香蕉霜冻害的保护效果,为进一步开发防霜剂和在实际生产实践中应用提供理论参考。
方法:(1)利用人工模拟气候室,先检测对香蕉产生冻害的临界温度;再将接种冰核细菌香蕉苗和无菌苗同时放到人工模拟气候室中,在相同的临界温度气候条件下,检测两组香蕉苗的冻害程度。
(2)采用经典抑菌圈法、药剂细菌的杀灭率和破坏冰核细菌冰核活性的等试验方法,初步筛选出一种有效的药剂;将所筛选出的药剂对香蕉进行药害试验,避免在施药后对香蕉产生药害。
(3)采用正交试验方法,优化防霜药剂SST溶剂配方;建立高效液相色谱法测定防霜药剂SST有效含量方法,进行防霜药剂稳定性试验。
(4)通过香蕉大田试验,检验在自然条件下防霜剂SST对香蕉叶片上冰核细菌数量的影响,以及对香蕉冻害的保护效果。
结果:(1)通过人工模拟气候室测定对香蕉产生冻害的临界温度为4℃,如果温度低于4℃,香蕉就会产生冻害;通过人工模拟霜冻试验证明了冰核细菌也是诱发和加重香蕉霜冻害的一个重要因素。
(2)经过3次抑菌、杀菌和破坏冰核细菌活性等试验,初步筛选出一种低毒、高效的防霜剂SST;SST稀释500倍及以上不会对香蕉产生药害作用;在人工模拟气候室条件下,防霜剂SST对香蕉的霜冻害具有很好的保护效果。
(3)采用正交试验方法,筛选出7.0%二甲基甲酰胺+30%TX-10+35%双链季胺为最佳的溶剂配方;经HPLC检测结果表明:SST在1.6～7.2μg·mL~(-1)的范围内,峰面积与进样量呈良好的线性关系(R=0.999 695)。方法的标准偏差(S)=0.0943,相对标准偏差(RSD)=0.0957%,平均回收率为97.88%,并具有良好的稳定性。
(4)大田香蕉园喷药试验表明,随着喷药次数的增加,香蕉上冰核细菌的数量随之下降,防霜剂SST在降低香蕉霜冻害方面具有明显的效果。
结论:试验表明冰核细菌也是诱发和加重香蕉霜冻害的一个主要因素,降低或杀灭香蕉上的冰核细菌,将是降低香蕉霜冻害的一种新型、方便有效的措施。
Objective: To verificate that INA bacteria is an important factor that induce and increase banana freezing injury through the artificial climate chamber simulation test; To obtain a kind of highly effective and lowly toxic anti-frost Pharmacy through INA bacteria inhibitory, killing, and destroying experiments. To work out the method of effective content testing and test its stability. To test the protective effects of the Pharmacy on bananas by field experiments and provide references for further practical utilizition of the Pharmacy.
Methods:(1) Identify the critical temperature at which banana freezing injury occurs using the artificial climate chamber simulation test and then detect the extent of damage of INA bacteria innoculated and non-innoculated banana seedlings at the same temperature.
(2) Use classic experiments such as the inhibition zone method, the killing rates test and the ice nucleation activity damage test to screen out a kind of effective pharmacy and test its phytotoxicity to make sure that it does not do damage to banana.
(3) To optimize pharmaceutical SST solvent formulations by orthogonal test; To work out the method of effective content testing using liquid chromatography experiments and test the stability of the pharmacy
(4) To detect the effect of the SST-cream on the quantity of ice nucleation bacteria and its protective effects against banana freezing damage through the artificial climate chamber simulation test.
Results:(1) Through artificial climate chamber tests the critical temperature at which banana freezing injury occurs was identified as 4℃. The INA bacteria was confirmed to be an important factor that induce and increase banana freezing damage.
(2) After three times of bacteriostatic, bactericidal and destruction of INA bacteria ice nucleation activity tests, the anti-cream SST was screened which has low toxicity and high efficiency; when diluted 500 times or above SST-cream did not have phytotoxicity activity against bananas; the SST-cream had significant protective effects against banana freezing injury in climatic chamber.
(3) By orthogonal test methods, it was found that 7.0% dimethyl formamide TX-30% + 10 + 35% quarter of double-stranded amine is the best solvent formula; the HPLC results showed that: in the 1.6 to 7.2μg / mL 1 area, the peak area and injection volume of SST showed a good linear relationship (R = 0.999 695), with precision of the standard deviation (S) = 0.0943, the relative standard deviation (RSD) = 0.0957% and the average recovery was 97.88%, and had a good stability.
(4) Field tests showed that the amount of INA bacteria decreased significantly following the spraying of the SST-cream. The SST-cream had significant effects in reducing the impact of the banana freezing damage.
Conclusion: It was confirmed that INA bacteria was a major factor that induce andincrease banana freezing damage. Reducing or killing INA bacteria on bananas wouldbe a convenient and effective alternative to control banana freezing damage.
引文
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