:256 Ethel McLennan : 



brown, followed by aniline gentian violet; later stages were also 

 treated in the same way. In addition, some of the later sections 

 were stained with congo red. These were lirst stained with aniline 

 gentian violet, followed by Gram's iodine water, and finally by 

 Congo red. This stain washes out very readily in the alcohols, so 

 it was found necessary to use a watery solution of congo red, and 

 to wash away excess with water, then to drain off as much of the 

 water as possible, and transfer immediately to clove oil. If the 

 sections be left on the water oven at a temperature of 45''-50°C., 

 the}^ will clear perfectly well in from 1-2 Ins., and they can then 

 be mounted iii balsam and piove to be quite permanent. 



7' he mature (jrain. 



My aim at first was to make a record of the grains of both Lolium 

 femuleiitiwi, and Lolium jyerenne examined, and to note the number 

 of fungus-containing and fungus-free seeds. 



After examining a large number of grains, I have been forced 

 to the conclusion that it is impossible to distinguish macroscopic- 

 ally grains containing the fungus from those devoid of it (if any). 

 The colour difference cited by Freeman cannot be regarded as a 

 'distinguishing feature. 



Nine' grains were chosen from a sample of Darnel obtained from 

 Northam, Western Australia. Of these 4 were very dark in 

 colour, 2 more or less intermediate, and 3 a pale straw yellow, but 

 all of the nine showed a dense hyphal layer situated between the 

 aleurone layer and the outer testa and pericarp. This is but a 

 single example of many similar series. xA.s the work proceeded it 

 became more and more evident that both colour and size of grain 

 were quite independent of the fungal constituent. 



When commencing this record hand sections were used, as it was 

 possible to handle a large number of fruits in a comparatively short 

 time, by this means. Sometimes these hand sections revealed a 

 grain apparently fungus-free — i.e., no definite layer of hyphae 

 could be seen in the usual position in the grain. These, w^hen ob- 

 tained, were frequently microtomed, and fine but distinct fungal 

 hyphae were found penetrating the scutellum, so it seemed impos- 

 sible to decide whether a particular grain was devoid of the fungus 

 unless serial sections were obtained sufficiently thin to enable these 

 fine threads to be demonstrated. Although hand sections are use- 

 ful in demonstrating the presence of the fungus, they cannot be 

 -accepted as evidence in regard to its absence. 



