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New method for testing condition of seeds
April 03, 2001Wageningen UR develops method for testing condition of seeds
Flow-cytometry, the technique for studying large numbers of individual cells in a fluid, can be combined with the use of special fluorescent colouring agents to form a valuable method of determining the health of seeds and seed consignments. This is shown in the thesis with which Brazilian Luiz G. Chitarra earned his doctorate on 28th March 2001 at Wageningen University. The technique will enable scientists to achieve results faster and more efficiently than ever before. Chitarra made his findings during his research at Plant Research International and Wageningen University.
Huge economic damage is caused every year by pathogenic bacteria living on and within seeds, which is why it is crucial to use pathogen-free seeds for sowing. The techniques currently being used to detect the existence and viability of such bacteria are generally complex and time consuming. Faster and more efficient techniques will therefore result in a better level of quality control and help avoid delays in the processing of seed consignments.
The dairy industry and the medical world use flow-cytometry to detect the presence and viability of various kinds of micro-organisms. Chitarra looked into the possible use of the technique for determining the presence, number and viability of the type of bacteria that cause diseases in plants. He studied Xanthomonas campestris pv. campestris, the pathogen that causes blackrot of crucifers in cabbage, and Clavibacter michiganensis subsp. michiganensis, the pathogen that causes bacterial wilt in the tomato.
In the case of the Xanthomonas bacteria, Chitarra proved that flow-cytometry is an efficient method for determining the number of pathogenic cells, even when there are other harmless bacteria present directly related to the pathogen. Chitarra used fluorescent antibodies that had been specially made for the study. These antibodies recognise the pathogen and attach themselves to its surface, colouring the cells. The closely related bacteria are not coloured by the antibodies. This method can therefore be used to determine the presence and number of pathogenic cells in seed samples.
In the case of the Clavibacter bacteria, Chitarra developed a method using flow-cytometry to determine which of the pathogens are still viable. He used a combination of two colouring agents. One of them colours only living bacteria while the other mainly attaches itself to dead bacteria. As a result, the flow-cytometer can accurately indicate whether or not a cell is still viable.
According to Chitarra, flow-cytometry will prove an excellent technique for detecting whether or not the micro-organisms in seeds are pathogenic and for determining whether they are still viable. As testing only takes a few hours the seeds need not be stored for long periods of time. Moreover, as a so-called 'high throughput technique', flow-cytometry is very useful for testing large amounts of samples and therefore more cost-effective than other options currently available.
Chitarra is planning to return to Brazil and continue his research into seed pathology, including pathogens that affect cotton and beans. He will co-operate in his work with Wageningen UR.
Wageningen University and Research Centre
The dairy industry and the medical world use flow-cytometry to detect the presence and viability of various kinds of micro-organisms. Chitarra looked into the possible use of the technique for determining the presence, number and viability of the type of bacteria that cause diseases in plants. He studied Xanthomonas campestris pv. campestris, the pathogen that causes blackrot of crucifers in cabbage, and Clavibacter michiganensis subsp. michiganensis, the pathogen that causes bacterial wilt in the tomato.
In the case of the Xanthomonas bacteria, Chitarra proved that flow-cytometry is an efficient method for determining the number of pathogenic cells, even when there are other harmless bacteria present directly related to the pathogen. Chitarra used fluorescent antibodies that had been specially made for the study. These antibodies recognise the pathogen and attach themselves to its surface, colouring the cells. The closely related bacteria are not coloured by the antibodies. This method can therefore be used to determine the presence and number of pathogenic cells in seed samples.
In the case of the Clavibacter bacteria, Chitarra developed a method using flow-cytometry to determine which of the pathogens are still viable. He used a combination of two colouring agents. One of them colours only living bacteria while the other mainly attaches itself to dead bacteria. As a result, the flow-cytometer can accurately indicate whether or not a cell is still viable.
According to Chitarra, flow-cytometry will prove an excellent technique for detecting whether or not the micro-organisms in seeds are pathogenic and for determining whether they are still viable. As testing only takes a few hours the seeds need not be stored for long periods of time. Moreover, as a so-called 'high throughput technique', flow-cytometry is very useful for testing large amounts of samples and therefore more cost-effective than other options currently available.
Chitarra is planning to return to Brazil and continue his research into seed pathology, including pathogens that affect cotton and beans. He will co-operate in his work with Wageningen UR.
Wageningen University and Research Centre
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