4 Stop es and Fujii, The nutritive relationa of the Burrounding tissues etc. 



Eood enters. Our vrork has impressed on us the remarkable 

 uniformity in the structure of the tbickened wall in all the Cycads, 

 Ginkgo, and Pinus in which the big pits are closed by a final 

 lamella which can oormally onlj allow soluble or semi-soluble i'ood 

 tu pass. 



All recenl workers appear to unite in turning linally to the 

 ouclei of the jackel eells as the factory of protein nourishmenl for 

 the egg cell, and do aol carry the question Enrther. Thus ignoring 

 the possible activities of the jackel cells themselves and their woik 

 as agents betweeo the original supply of irarious i'ood substances 

 and the growing egg in whieh the food is required in a t'orni 

 available for immediate utilization. 



Materials and their treatment. 



In the conrse of our investigations up to the presenl vre 



have examined the following species: Cycas Beddomei, C. circhtal.is, 

 C. Normanbyana, C. revoluta, and C. sp.?, Zamia ßoridana, Z. integri- 

 folia, Z. muricata and Z. sp.?, Ceratozarnia fusco-wridis, C. mexicana, 

 C. Miqueliana, Macrozamia J'reissii, M. spiralis, Eneephcdartos Hilde- 

 brandtii, K. horridus, E. Lehmanni and E. sp.?, Dioon edule, Stangeria 

 schizodon, Ginkgo biloba, Pinus Cembra, P. montana, P. Pinea and 

 P. sylvestris. 



In this first part of our work we have used for rlxing 

 Flemrnings strong Solution diluted witli an equal volume of water; 

 Alcohol acetic; Chrom-acetic; and various strengths of Alcohol 

 alone. In the case of the Cycads the best preservation of the 

 structure of the egg cytoplasm without shrinkage was observed 

 after usiiig 90 °/ alcohol, lixing the whole seed even when the 

 stone layers hacl considerably hardened. For digestion experiments 

 with pepsin also 90 °/ alcohol proved to be the best fixative. 

 AN'ith Flemming's fixative we found that a very strong reducing 

 substance present in the tissues caused. excessive deposit of reduced 

 osniiuin. and the results were not always satisfactory. With Pinus, 

 particnlarly P. Cembra, where the endosperm is very large, we got 

 excellent preservation of the structure of the egg cytoplasm by 

 separating the endosperm from the integiimcnts and nncellus, 

 fixing in 30 ° Alcohol and then slowly transferring to higher 

 percentages of alcohol. 



We also always examined fresh material whenever it was 

 possible, for many of the substances which are important in the 

 process of nutrition such as oxydases, sugars etc., cannot be dealt 

 with in fixed material. 



For paraffin embedding we used Cedar oil chiefly (sometimes 

 Chloroform) between the Alisolute Alcohol and paraffin. Microtome 

 series were stained with Flemrnings triple stain, acetic methyl green 

 (with or without the addition of Sodium sulphate) Congo red, 

 ruthenium red. and other stains. or treated with acpieous Solution 

 of iodine, alcoholic Solution of iodine in potassium iodide, Millon's 

 reagent, or Chlorzinc Iodine as the case demanded. Microtome 



