FGSC #5176

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Reporting Genes: sn cr-1;acr-2 trp-1 dow

Species: crassa

Allele: C136 B123 KH5(r) 10575 P616

Alternate Strain Number: M1863

Depositor: EK

Linkage Group: I IR;III IIIR IIIR

Mating Type: a

Species Number: 10

Deposit Sheet ↗

Genes

Locus	Cultural Requirements	Link Group	Type
acr-2	III. Linked to thi-4 (0/286). Left of sc (3 to 6%) and spg (1 to 11%) (498, 816). acr-2 has been shown left of the centromere on published maps but without direct evidence. acr-2and trp-1 (on IIIR) cosegregated at the second division in 1 of 13 asci (H.B. Howe, Jr., personal communication), which would favor a right arm location. Resistant to acriflavine (494, 495); also resistant to 3-amino-1,2,4-triazole (seven alleles tested) (494). Resistance is probably dominant (heterokaryon tests) (498). Not resistant to malachite green. An excellent stable marker, fully fertile, with unambiguous scoring. Sizable inocula should be used to avoid false-negative tests. Use acriflavine at 50 µg /ml in minimal agar medium (816) (higher concentrations may be used) and aminotriazole at 0.5 mg/ml; both added before autoclaving.	III	B
cr-1	IR. Right of ace-7 (1 to 3%) and nic-2 (4 to 7%). Left of cys-9 (3%) and un-1(5%) (721, 816). Included in duplications from T(4540), which do not include cr-2 or cr-3(PB). (610) Rapid conidiation close to surface of agar. Produces very short conidiophores, bearing conidia in tight clusters (610, 611). Photographs (533, 634). Recessive. Deficient in adenylate cyclase (1066); has little or no endogenous adenosine 3',5'-phosphate (1065, 779). Abnormal morphology partially corrected by exogenous adenosine 3',5'-phosphate (891, 892, 1065, 1066). Guanosine 3',5'-phosphate also stimulates mycelial elongation (892). Cyclic nucteotide levels differ in mycelia and conidia (891, 892). NAD(P) glycohydrolase is overproduced and excreted; this is normalized by adenosine 3',5'-phosphate (533). Induction and localization of p-glucosidase is altered; induction is normalized by adenosine 3',5'-phosphate (906). Inability to use glycerol and certain other carbon sources is also overcome by adenosine 3',5'phosphate (598, 1067). Phosphodiesterase inhibitors do not counteract the morphological effect of cr-1 (892). Increased lactate dehydrogenase activity (92). Used to determine what functions are controlled by adenosine 3',5'-phosphate (779). Used to study adenosine 3',5'-phosphate binding protein (1082). Strains carrying the various alleles vary in growth habit (B123 strains are flat, restricted; allele L strains are spreading, but morphology may vary on different media). Modifier mutations which alter morphology and the ability of cr-1 to use glycerol occur frequently (383, 905). Crosses homozygous for allele B123 exude intact linear asci (634). Double mutants sn cr and cr rg form small conidiating colonies suitable for replica plating with velvet (182, 634, 796, 932, 1020). The triple mutant sn cr;csp-2 can be overlayered (744; photograph 747). The single mutant (B123) can be replicated by using a needle replicator (634). Scorability and viability are good. Excellent as a marker. Carotenoids formed normally. cr-1 ascospores may require longer to mature than cr+ ascospores. Allele CE4-11-67 called con(716, 717).	IR	B
dow	IIIR. Right of ty-1 (21%) and un-17 (23%); linked to erg-3 (10%) and acu-7(0/72) (816).Soft, matty growth, conidiating and covering slants (816). An excellent marker: good viability, fertility, and scorability.	IIIR	B
sn	I. Right of T(39311) and arg-3 (1 to 6%). Left of T(AR173) and his-2 (<1 to 12%) (174, 808). (687) Spreading colonial growth with good conidiation. Linear growth is less than 1/10 that of the wild type (19). Detectable immediately after ascospore germination by hyphal patterns which suggested the name (688). Abnormal microfilaments (19). Contains actin-like protein (20). Said not to exhibit cytoplasmic streaming (18). Meiosis and ascospore formation are normal in homozygous sn x sn crosses (N.B. Raju, personal communication). Good female fertility. Morphology similar to that of sp, cum, and cot-4mutants (at 25 C) (PB). Used to study development of crystalline inclusions (17). The cr sn double mutant grows as small, discrete, conidiating colonies suitable for velvet replication. The double mutant cr snresembles the rg cr double mutant phenotypically and has the advantage of fertility in homozygous crosses (796); for example of application, see reference 180.	IR	B
trp-1	IIIR. Between ad-2 (1 to 7%) and ro-2 (2 to 12%) (11, 219, 812). Linked to fpr-3 (< 1%) (550). (504). Uses tryptophan or indole (1060); strains carrying some alleles can also use anthranilate; others cannot (4). trp-1+ and trp-2+ gene products together form an enzyme aggregate with three activities: anthranilate synthetase, phosphoribosyl-anthranilate isomerase, and indoleglycerol-phosphate synthetase (181, 260) (Fig. 11). trp-1 codes for the beta subunit of the aggregate (546); it specifies phosphoribosyl-anthranilate isomerase, indoleglycerol-phosphate synthetase, and collaboratively the glutamine amino transferase activity of anthranilate synthetase (29, 181, 502). Strains carrying different alleles differ in lacking one or more of the three activities, e.g., trp-1 (allele 15) lacks all three activities; trp-1 (20) lacks only phosphoribosyl- anthranilate isomerase, trp-1C (1) lacks only anthranilate synthetase, trp-1 (25) lacks both phosphoribosyl-anthranilate isomerase and indoleglycerol-phosphate synthetase, etc. (259). (To avoid confusion, note that in reference 259 and related papers, the same "allele number" may be used for a trp-2 mutation, a trp-1 mutation [non-anthranilate-utilizing], and a trp-1Cmutation [anthranilate utilizing]; mutations of the last class are listed by FGSC as trp-1 with the allele number prefixed by C.) Strains carrying different alleles differ in their ability to form aggregates (181, 259). Association between trp-1 and trp-2products is essential for glutamine-dependent anthranilate synthetase activity but not the other two activities (181). The trp-1 gene has been cloned (545, 925), sequenced (925), and reintroduced into Neurospora by transformation (925). It is only partially expressed in E. coli. Fine-structure maps (10, 259). Complementation maps (10, 163). Reviewed as example of gene fusion (218). Nonsense allele used to demonstrate restoration of normal enzyme aggregate by supersuppressors (183). Alleles that accumulate anthranilate are scorable by blue fluorescence under long-wave UV after 2 to 5 days of growth on minimal medium plus indole (10 µg /ml), 34°C (814, 816). Aging cultures may produce brown pigment; blue fluorescence disappears as pigment forms.	IIIR	B