Professional Documents
Culture Documents
Nitrogen Requirements and Deficiencies of Thaxter: Piiytophthora
Nitrogen Requirements and Deficiencies of Thaxter: Piiytophthora
DEFICIENCIES O F P I I Y T O P H T H O R A P H A S E O L I
THAXTER
BY R. K. SAKSENAAND K. S. BHARGAVA
(Botany Department, University of Allahabad)
Received July 15, 1943
(Communicated by Prof. Shri Ranjan, D.sc., F.A.SC.)
IT has been well established that the source of nitrogen, in the cultural
nutrition of fungi, is an important limiting factor (Robbins, 1937, Leonian and
Lilly, 1938), but for some even the proper source of nitrogen often may be
ineffective without the presence of growth supplements. Robbins (1938),
who studied the nutrition of some species of Phytophthora, found that they
apparently required for their growth an external supply of thiamin. Leonian
(1925, p. 448) has reported that Phytophthora phaseoli grows very slowly on
the two media used by him. One contained malt extract (dry), KH2PO4,
MgSO, and Bacto agar, while the other was made up of nucleinic acid,
KH~PO4, MgSO4, dextrose and Bacto agar. The latter medium was used
without agar also. When he added a few c.c. of lima bean infusion to the
media, the fungus showed rapid growth. His results indicate that P.
phaseoli suffers from a deficiency of some growth supplements not amply
represented in the original media used by him.
The present paper deals with the nitrogen requirements and vitamin
deficiencies of Phytophthora phaseoli Thaxter.
For a comprehensive review on these subjects the reader is referred to
Robbins (1937) and Robbins and Kavanagh (1942).
Material and Methods
The culture of Phytophthora phaseoli Thaxter was obtained from
Centraal Bureau voor Shimmelcultures, Baarn, Holland.
The methods and technique employed in this investigation were essentially
the same as described in previous papers (Saksena, 1941 a and b; Saksena
and Bhargava, 1941).
Unless otherwise indicated the fungus was grown in 150 ml. Erlenmeyer
Pyrex flasks containing 15 c.c. of a basal medium, which will afterwards be
referred to as medium M, consisting of 0.5 gm. each of KH~PO4, MgCI~"
6HzO and K~SO4, 5.0 gm. of pure dextrose (dextrosol of Corn Products Co.)
45
B3
46 R . K . Saksena and K. S. Bhargava
• Indicates absence o f g r o w t h .
Series VI b.--To know exactly whether the ammonium ion or the nitrate
ion is responsible for the toxicity Lconian's medium used in Series V I a was
supplemented with thiamin (5 units per 25 c.c.), and NH4CI, (NH4)s SO 4 and
NaNO3 were added to it singly in 0"2~o concentration. The fungus showed
growth on Leonian's medium supplemented with thiamin and on the medium
containing NaNO3 while it did not grow on those to which NH4CI or
(NH4)~ SO4 had been added.
Discussion and Conclusions
Earlier work of Leonian (1925, 1930) on the nutrition of the genus
Phytophthora furnishes little information on the nutritional requirements of
Phytophthora phaseoli. The inability of P. phaseoli to grow on nutrient
media containing inorganic nitrogen (Series I) clearly indicates that probably
it is one of those fungi which do not assimilate ammonia or nitrate but require
some organic nitrogen. The positive reaction which the organism shows
Nitrogen Requirements & Vitamin De/qciencies o f P. phaseoli Thaxter 49
with d-alanine and peptone .justifies its being placed in the ' organic nitrogen
organisms' classified and placed separately by Robbins (1937). Kincaid
(Robbins 1937, p. 244) found that Blepharospora eambivora, Diplodia Zeee,
Endomyces Magnusii, Phyeomyees nitens, thirteen species of Phytophthora,
Spheeronema fimbriatum did not grow with either nitrate or ammonia as
the source of nitrogen and dextrose as the source of carbon but grew with
peptone. Similarly Euglena deses and E. pisei-formis do not assimilate
nitrate or ammonia but require amino acids or peptone (Dusi, 1933).
But the relation of organisms to organic nitrogen is generally complicated
by the possible contamination of organic nitrogen compounds with the
presence of accessory growth factors, which are not generally found in pure
inorganic salts. In the present case, d-alanine, peptone, hydrolysed peptone
and butter milk are found to contain traces of thiamin or its intermediates
(Series IIO. Since lentil extract (Hawker, 1936, p. 703) and yeast extract
(Leonian and Lilly, 1938) in addition to some of the known vitamins
contain amino acids and other organic nitrogen compounds, it is clear that
they supply the necessary source of organic nitrogen in addition to thiamin
(Series V).
That thiamin is also essential for the growth of P. phaseoli is shown by
the fact that it fails to grow on medium M containing purified d-alanine but
shows good growth in the presence of d-alanine (Series II), which has been
demonstrated to contain thiamin 6r its intermediates (Series lII).
This conclusion is also supported by the experiment carried under Series IV
where the addition of thiamin to medium M containing purified d-alanine
or purified casein induces the growth of the organism. It may be noted here
that the fungus grows well on medium M containing lime bean infusion
(Series V). This shows that lima bean infusion is a source both of a suitable
organic nitrogen and growth supplement for the organism.
Since peptone is a complex mixture containing minerals, amino acids,
several vitamins as well as other nitrogen compounds, no important conclu-
sion as regards nitrogen requirements of the fungus can be derived from
its use. Similar is the case with lentil extract, yeast extract and lima bean
infusion. The most important result is the presence of growth on medium
M containing purified d-alanine supplemented with thiamin (Series IV).
This shows that P. phaseoli requires a special amino acid as source of nitro-
gen for its growth and that it suffers from thiamin deficiency. Both amino
acids and thiamin are considered necessary by Leonian and Lilly (1938) for
Coprinus lagopus, Cha~tocladium Brefeldii, Nyctalis asterophora, Pilaira
moreaui and Pleurotus corticatus also.
50 R . K . Saksena and K. S. Bhargava